Methods of Regulating Oxidoreductase Activity for Treatment of Inflammation and Age-Related Disorders

ABSTRACT

In some aspects, a method of regulating oxidoreductase activity for treating an inflammation or age-related disorder involves administering to an individual isomyosmine or a pharmaceutically acceptable salt thereof. In other aspects, a method of treating oxidative stress associated with an inflammation or age-related disorder involves administering to an individual a therapeutically effective amount of isomyosmine or a pharmaceutically acceptable salt thereof. In other aspects, isomyosmine or a pharmaceutically acceptable salt thereof may be administered to an individual for the treatment of infectious or parasitic diseases or various other disorders.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of U.S. application Ser. No.16/680,677, filed Nov. 12, 2019, which is a continuation of U.S.application Ser. No. 15/719,875, filed Sep. 29, 2017, now U.S. Pat. No.10,517,856 B2, which is a continuation-in-part of PCT/US16/25126, filedMar. 31, 2016, which in turn claims priority under 35 U.S.C. § 119(e) toU.S. App. No. 62/140,618, filed Mar. 31, 2015, the disclosures of eachof which are hereby incorporated by reference in their entireties.

BACKGROUND

Oxidoreductases are a class of enzymes that catalyze the transfer ofelectrons from reductants (electron donors) to oxidants (electronacceptors). This type of reaction is also known as an oxidoreductionreaction. The reaction generally follows the following scheme where A isthe reductant and B is the oxidant:

A ⁻ +B→A+B ⁻

Oxidoreductases can be oxidases or dehydrogenases. Oxidases are enzymesinvolved when molecular oxygen acts as an acceptor of hydrogen orelectrons. Dehydrogenases are enzymes that oxidize a substrate bytransferring hydrogen to an acceptor that is either NAD⁺/NADP⁺ or aflavin enzyme. Other oxidoreductases include peroxidases, hydroxylases,oxygenases, and reductases. Peroxidases are localized in peroxisomes,and catalyzes the reduction of hydrogen peroxide. Hydroxylases addhydroxyl groups to its substrates. Oxygenases incorporate oxygen frommolecular oxygen into organic substrates. Reductases catalyzereductions, in most cases reductases can act like an oxidases.

Oxidoreductase enzymes play an important role in both aerobic andanaerobic metabolism. They can be found in glycolysis, TCA cycle,oxidative phosphorylation, and in amino acid metabolism. In glycolysis,the enzyme glyceraldehydes-3-phosphate dehydrogenase catalyzes thereduction of NAD⁺ to NADH. In order to maintain the re-dox state of thecell, this NADH must be re-oxidized to NAD⁺, which occurs in theoxidative phosphorylation pathway. Additional NADH molecules aregenerated in the TCA cycle. The product of glycolysis, pyruvate entersthe TCA cycle in the form of acetyl-CoA. During anaerobic glycolysis,the oxidation of NADH occurs through the reduction of pyruvate tolactate. The lactate is then oxidized to pyruvate in muscle and livercells, and the pyruvate is further oxidized in the TCA cycle. All twentyof the amino acids, except leucine and lysine, can be degraded to TCAcycle intermediates. This allows the carbon skeletons of the amino acidsto be converted into oxaloacetate and subsequently into pyruvate. Thegluconeogenic pathway can then utilize the pyruvate formed.

The aging process is known to be associated with increased oxidativestress, that is with accumulation in the cells of reactive oxygenspecies (ROS), which are mainly represented by superoxide anion,hydrogen peroxide, and hydroxyl radical. ROS accumulation damagesnumerous types of biological molecules, such as proteins, lipids, or DNA(FIG. 3), with proteins being the most dominant target. Tanase et al.,“Role of Carbonyl Modifications on Aging-Associated ProteinAggregation,” Sci Rep 6, 19311 (2016). ROS induce post-translationalmodifications of proteins including glycation, glycoxidation,lipoxidation, and carbonylation. Dalle-Donne, “Protein carbonyl groupsas biomarkers of oxidative stress,” Clinica Chimica Acta. Vol. 329,Issues 1-2, March 2003, pp. 23-38. Protein carbonyl content is the mostcommonly used indicator of protein oxidation. Accumulation of proteincarbonyls is seen in several human inflammatory and age-related diseasesincluding Alzheimer's disease, diabetes, inflammatory bowel disease andarthritis. Berlett et al., “Protein oxidation in aging, disease, andoxidative stress,” J Biol Chem 1997; 272:20313-6; Uchida, “Role ofreactive aldehyde in cardiovascular diseases,” Free Radic Biol Med 2000;28:1685-96; Stadtman et al., “Reactive oxygen-mediated protein oxidationin aging and disease,” Chem Res Toxicol 1997; 10:485-94.

The use of protein carbonyl groups as a biomarker of oxidative stressmay be more appealing than the measurement of other oxidation productsbecause of the relative early formation and stability of carbonylatedproteins. Dalle-Donne, supra. A highly sensitive assay was developed forthe detection of protein carbonyls that involves the reaction of thecarbonyl group with 2,4-dinitrophenylhydrazine (DNPH), which leads tothe formation of a stable 2,4-dinitrophenyl (DNP) hydrazone product. Themeasurement of protein carbonyls following their covalent reaction withDNPH has become the most widely utilized measure of protein oxidation inseveral human diseases. Levine et al., “Determination of carbonylcontent in oxidatively modified proteins,” Methods Enzymol 1990; 186:464-78.

An enzyme-linked immune-adsorbent assay (ELISA) method was developedusing an anti-DNP antibody for measuring total protein carbonyl groupsthat is highly sensitive, reproducible, and correlates directly with theclassical colorimetric assay. Buss et al., “Protein carbonyl measurementby a sensitive ELISA method,” Free Radic Biol Med 1997; 23(3):361-6. TheELISA test has the important advantage that requires only microgramamounts. Therefore, ELISA has a wide application for measuring proteinoxidation both experimentally and clinically in situations where onlylimited amounts of protein are available for analysis.

Xanthine oxidoreductase (XOR) catalyzes the final two reactions thatlead to uric acid formation (FIG. 4). XOR is a complexmolibdo-flavo-enzyme present in two different functional forms:dehydrogenase and xanthine oxidase (XO). Della Corte et al., “Propertiesof the xanthine oxidase from human liver,” Biochim Biophys Acta., 1969,191(1), pp. 164-166. Under physiological conditions, it is mainly foundin the dehydrogenase form, with the highest levels found in intestineand liver, but during inflammatory conditions, stress phenomena andaging, it is easily converted into XO by oxidation of the sulfhydrylresidues or by proteolysis and release into the circulation. Kelley etal., “Hydrogen peroxide is the major oxidant product of xanthineoxidase,” Free Radic Biol. Med., 2010, 48(4), 493-8; Glantzounis et al.,“Uric acid and oxidative stress,” Curr Pharm Des., 2005, 11(32):4145-51.XO is a critical source of reactive oxygen species (ROS). Fully reducedXO contains a total of six electrons (two each at the flavin andmolybdenum sites, and one each at the two iron-sulfur centers) and itsre-oxidation involves electron transfer to oxygen molecules whichgenerates two H₂O₂ and two O₂ ⁻ species for every full reduced XOmolecule (FIG. 4); George et al., “Role of urate, xanthine oxidase andthe effects of allopurinol in vascular oxidative stress,” Vasc HealthRisk Manag., 2009, 5(1), 265-72.

SUMMARY

In some aspects, a method of regulating oxidoreductase activity fortreating an inflammation or age-related disorder comprises administeringto an individual in need thereof a therapeutically effective amount ofisomyosmine or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable vehicle therefor. In some examples, theinflammation-related disorder is a parasitic infection or disease suchas malaria. In other examples, the inflammation-related disorder is abacterial infection or disease such as Lyme disease.

In other examples, a method of treating oxidative stress associated withan inflammation or age-related disorder comprises administering to anindividual in need thereof a therapeutically effective amount ofisomyosmine or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable vehicle therefor.

In yet other examples, a method of reducing oxidative stress in anindividual suffering from an inflammation or age-related disordercomprises administering to the individual a therapeutically effectiveamount of isomyosmine or a pharmaceutically acceptable salt thereof, anda pharmaceutically acceptable vehicle therefor.

In other aspects, a method of treating a disorder comprisesadministering to an individual in need thereof a therapeuticallyeffective amount of isomyosmine or a pharmaceutically acceptable saltthereof, where the disorder is selected from the group consisting ofinfectious or parasitic diseases; neoplasms; diseases of the blood orblood-forming organs; diseases of the immune system; endocrine;nutritional or metabolic diseases; mental, behavioral orneurodevelopmental disorders; sleep-wake disorders; diseases of thenervous system; diseases of the visual system; diseases of the ear ormastoid process; diseases of the circulatory system; diseases of therespiratory system; diseases of the digestive system; diseases of theskin; diseases of the musculoskeletal system or connective tissue;diseases of the genitourinary system; conditions related to sexualhealth; pregnancy, childbirth or the puerperium; conditions originatingin the perinatal period; developmental anomalies; and injury, poisoningor other consequences of external causes. In one example, a behavioralor neurodevelopmental disorder is anxiety. In another example, abehavioral or neurodevelopmental disorder is depression.

BRIEF DESCRIPTION OF THE DRAWINGS

A more complete understanding of the present invention and certainadvantages thereof may be acquired by referring to the followingdetailed description in consideration with the accompanying drawings, inwhich:

FIGS. 1A-1B show hydrogen sulfide relative quantities in mice with EAE,treated either with a nitrate reductase inhibitor or a control;

FIGS. 2A-2C illustrate the ability of isomyosmine to directly inhibitnitrate reductase in a cell-free setting with low substrate (FIG. 2A),medium substrate (FIG. 2B), and high substrate (FIG. 2C);

FIG. 3 illustrates carbonylation of biomolecules and its cytotoxiceffects; and

FIG. 4 illustrates xanthine oxidase catalyzing the final two reactionsin the biochemical chain that lead to uric acid formation, particularlythe conversion of hypoxanthine to xanthine and xanthine to uric acidwhich is responsible for ROS generation.

DETAILED DESCRIPTION

Aspects of the present specification disclose, in part, a pharmaceuticalcomposition. As used herein, the term “pharmaceutically acceptable”means any molecular entity or composition that does not produce anadverse, allergic or other untoward or unwanted reaction whenadministered to an individual. As used herein, the term“pharmaceutically acceptable composition” is synonymous with“pharmaceutical composition” and means a therapeutically effectiveconcentration of an active ingredient, such as any of the therapeuticcompounds disclosed herein. A pharmaceutical composition is useful formedical and veterinary applications. A pharmaceutical composition may beadministered to an individual alone, or in combination with othersupplementary active ingredients, agents, drugs or hormones.

A pharmaceutical composition may include a pharmaceutically acceptablecarrier that facilitates processing of an active ingredient intopharmaceutically acceptable compositions. As used herein, the term“pharmacologically acceptable carrier” is synonymous with“pharmacological carrier” and means any carrier that has substantiallyno long term or permanent detrimental effect when administered andencompasses terms such as “pharmacologically acceptable vehicle,”“stabilizer,” “diluent,” “additive,” “auxiliary” or “excipient.” Such acarrier generally is mixed with an active compound or permitted todilute or enclose the active compound and can be a solid, semi-solid, orliquid agent. It is understood that the active ingredients can besoluble or can be delivered as a suspension in the desired carrier ordiluent. Any of a variety of pharmaceutically acceptable carriers can beused including, without limitation, aqueous media such as, e.g., water,saline, glycine, hyaluronic acid and the like; solid carriers such as,e.g., mannitol, lactose, starch, magnesium stearate, sodium saccharin,talcum, cellulose, glucose, sucrose, magnesium carbonate, and the like;solvents; dispersion media; coatings; antibacterial and antifungalagents; isotonic and absorption delaying agents; or any other inactiveingredient. Selection of a pharmacologically acceptable carrier candepend on the mode of administration. Except insofar as anypharmacologically acceptable carrier is incompatible with the activeingredient, its use in pharmaceutically acceptable compositions iscontemplated. Non-limiting examples of specific uses of suchpharmaceutical carriers can be found in Pharmaceutical Dosage Forms andDrug Delivery Systems (Howard C. Ansel et al., eds., Lippincott Williams& Wilkins Publishers, 7th ed. 1999); REMINGTON: THE SCIENCE AND PRACTICEOF PHARMACY (Alfonso R. Gennaro ed., Lippincott, Williams & Wilkins,20th ed. 2000); Goodman & Gilman's The Pharmacological Basis ofTherapeutics (Joel G. Hardman et al., eds., McGraw-Hill Professional,10th ed. 2001); and Handbook of Pharmaceutical Excipients (Raymond C.Rowe et al., APhA Publications, 4th edition 2003). These protocols areroutine procedures and any modifications are well within the scope ofone skilled in the art and from the teaching herein. Formulationscontaining isomyosmine or a dietary acceptable salt thereof similarlymay be formulated as dietary supplements or nutraceuticals usingmaterials and techniques well known to persons skilled in the art.“Dietary acceptable” as used herein is synonymous with “pharmaceuticallyacceptable” but refers to dietary supplement compositions rather thanpharmaceutical compositions per se.

An “age-related disorder,” as used herein, refers to a disease that ismost often seen with increasing frequency with increasing senescence.Age-related disorders essentially are complications arising fromsenescence or, stated differently, diseases associated with the elderly.Non-limiting examples of age-related diseases include atherosclerosisand cardiovascular disease, cancer, arthritis, cataracts, osteoporosis,type 2 diabetes, hypertension, and Alzheimer's disease. The incidence ofall of these diseases increases exponentially with age.

In some aspects, isomyosmine or a pharmaceutically acceptable saltthereof is administered as an oxidoreductase inhibitor at atherapeutically effective dose for modulating hydrogen sulfide andnitrite levels in an individual, which may be effective for treatinginflammation and age-related disorders. Such methods also may beeffective for promoting or supporting health in an individual, such asby promoting overall wellness, maintaining cell integrity, supportingheart health, supporting a healthy immune system, supporting colonhealth, supporting prostrate health, supporting brain health, supportingliver health, supporting nervous system health, supporting restfulsleep, supporting glucose metabolism, supporting the circulatory system,supporting eye health, providing key nutrients to support glucosemetabolism, providing key nutrients to support a normal glucoseutilization and insulin support, supporting a health inflammatoryresponse, supporting healthy blood sugar levels, protecting cells andsupporting healthy circulation, supporting normal healthy blood pressurelevels, enhancing blood vessel dilation, and/or supporting healthy blooddevelopment.

The chemical reactions catalyzed by oxidoreductases (A⁻+B→A+B⁻) areprone to lead to chronic inflammation and senescence in individuals. Byadministering an oxidoreductase inhibitor as described herein, theformation of excessive oxidants (B) may be reduced, leading to improvedchronic inflammatory states and senotherapy.

Oxidative stress caused by reactive species, including reactive oxygenspecies, reactive nitrogen species, and unbound, adventitious metal ions(e.g., iron [Fe] and copper [Cu]), has been identified as an underlyingcause of various neurodegenerative diseases, including depression andanxiety-related disorders. Such reactive species are an inevitableby-product of cellular respiration and other metabolic processes thatmay cause the oxidation of lipids, nucleic acids, and proteins. Themanifestation of anxiety in numerous psychiatric disorders, such asgeneralized anxiety disorder, depressive disorder, panic disorder,phobia, obsessive-compulsive disorder, and posttraumatic stressdisorder, highlights the importance of studying the underlying biologyof these disorders to gain a better understanding of the disease and toidentify common biomarkers for these disorders. The expression ofglutathione reductase 1 and glyoxalase 1, which are genes involved inanti-oxidative metabolism, were reported to be correlated withanxiety-related phenotypes. See Hassan et al., “Association of OxidativeStress to the Genesis of Anxiety: Implications for Possible TherapeuticInterventions,” Current Neuropharmacology, 12, 120-139 (2014).

Isomyosmine (3-(3,4-dihydro-2H-pyrrol-2-yl)-pyridine) is a nicotinerelated alkaloid present in solanecea plants containing nicotine.Isomyosmine may be prepared synthetically using known techniques, andalso is commercially available from several chemical suppliers.Isomyosmine has two optical isomers (+/−) owing to an asymmetric carbonatom within its pyrrole ring that joins to the pyridine ring. Unlessotherwise clear from context, the term “isomyosmine,” as used herein, isinclusive of enantiomeric mixtures (+/−) including racemic mixtures, aswell as isolated forms of one or the other enantiomer.

Unless otherwise clear from context, “isomyosmine” as used herein refersto both salt and non-salt forms of isomyosmine. Non-limiting examples ofpossible salts are described in P. H. Stahl et al., Handbook ofPharmaceutical Salts: Properties, Selection and Use,Weinheim/Zürich:Wiley-VCHNHCA, 2002, including salts of1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid,2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoicacid, 4-aminosalicylic acid, acetic acid, adipic acid, ascorbic acid(L), aspartic acid (L), benzenesulfonic acid, benzoic acid, camphoricacid (+), camphor-10-sulfonic acid (+), capric acid (decanoic acid),caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonicacid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid,ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaricacid, galactaric acid, gentisic acid, glucoheptonic acid (D), gluconicacid (D), glucuronic acid (D), glutamic acid, glutaric acid,glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid,hydrochloric acid, isobutyric acid, lactic acid (DL), lactobionic acid,lauric acid, maleic acid, malic acid (−L), malonic acid, mandelic acid(DL), methanesulfonic acid, naphthalene-1,5-disulfonic acid,naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid,oxalic acid, palmitic acid, pamoic acid, phosphoric acid, proprionicacid, pyroglutamic acid (−L), salicylic acid, sebacic acid, stearicacid, succinic acid, sulfuric acid, tartaric acid (+L), thiocyanic acid,toluenesulfonic acid (p), and undecylenic acid.

As an alternative to preparing isomyosmine synthetically, isomyosminecan be obtained by extraction from tobacco or other sources in which itoccurs naturally. For example, a tobacco extract may be prepared fromcured tobacco stems, lamina, or both. In the extraction process, curedtobacco material is extracted with a solvent, typically water, ethanol,steam, or carbon dioxide. The resulting solution contains the solublecomponents of the tobacco, including isomyosmine. Isomyosmine may bepurified from the other components of the tobacco using suitabletechniques such as liquid chromatography.

In pharmaceutical applications, an isolated form of isomyosminegenerally is used. An “isolated form of isomyosmine,” as used herein,refers to isomyosmine that either has been prepared synthetically or hasbeen substantially separated from natural materials in which it occurs.The isolated form of isomyosmine should have a very high purity(including enantiomeric purity in the case where an enantiomer is used).In the case of synthetic isomyosmine, for example, purity refers to theratio of the weight of isomyosmine to the weight of the end reactionproduct. In the case of isolating isomyosmine from native material, forexample, purity refers to the ratio of the weight of isomyosmine to thetotal weight of the isomyosmine-containing extract. Usually, the levelof purity is at least about 95%, more usually at least about 96%, about97%, about 98%, or higher. For example, the level of purity may be about98.5%, 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%,99.9%, or higher.

A pharmaceutical composition alternatively may contain one or moreoxidoreductase inhibitors and/or nitrate reductase inhibitors other thanisomyosmine. Suitable compounds having the ability to inhibit thereduction of nitrate (NO₃ ⁻) to nitrite (NO₂ ⁻) are known to personsskilled in the art and/or may be readily identified using an appropriateassay to test for this property. For example, a compound may be assayedtogether with nitrate reductase enzyme in vitro to determine whether andto what extent the compound has the ability to inhibit the reduction ofnitrate to nitrite. See Example 2 below.

A pharmaceutical composition can optionally include, without limitation,other pharmaceutically acceptable components (or pharmaceuticalcomponents), including, without limitation, buffers, preservatives,tonicity adjusters, salts, antioxidants, osmolality adjusting agents,physiological substances, pharmacological substances, bulking agents,emulsifying agents, wetting agents, sweetening or flavoring agents, andthe like. Various buffers and means for adjusting pH can be used toprepare a pharmaceutical composition disclosed herein, provided that theresulting preparation is pharmaceutically acceptable. Such buffersinclude, without limitation, acetate buffers, citrate buffers, phosphatebuffers, neutral buffered saline, phosphate buffered saline and boratebuffers. It is understood that acids or bases can be used to adjust thepH of a composition as needed. Pharmaceutically acceptable antioxidantsinclude, without limitation, sodium metabisulfite, sodium thiosulfate,acetylcysteine, butylated hydroxyanisole and butylated hydroxytoluene.Useful preservatives include, without limitation, benzalkonium chloride,chlorobutanol, thimerosal, phenylmercuric acetate, phenylmercuricnitrate, a stabilized oxy chloro composition and chelants, such as,e.g., DTPA or DTPA-bisamide, calcium DTPA, and CaNaDTPA-bisamide.Tonicity adjustors useful in a pharmaceutical composition include,without limitation, salts such as, e.g., sodium chloride, potassiumchloride, mannitol or glycerin and other pharmaceutically acceptabletonicity adjustor. The pharmaceutical composition may be provided as asalt and can be formed with many acids, including but not limited to,hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, etc.Salts tend to be more soluble in aqueous or other protonic solvents thanare the corresponding free base forms. It is understood that these andother substances known in the art of pharmacology can be included in apharmaceutical composition.

In some aspects, a pharmaceutical composition comprises at least oneoxidoreductase inhibitor and a pharmaceutically acceptable adjuvant. Inanother embodiment, a pharmaceutical composition comprises at least oneoxidoreductase inhibitor, a pharmaceutically acceptable solvent, and apharmaceutically acceptable adjuvant. In some aspects, a pharmaceuticalcomposition may further comprise a pharmaceutically acceptablestabilizing agent. In some aspects, a pharmaceutical composition mayfurther comprise a pharmaceutically acceptable carrier, apharmaceutically acceptable component, or both.

Compositions may contain at least one oxidoreductase inhibitor, alone orwith other therapeutic compound(s). A therapeutic compound is a compoundthat provides pharmacological activity or other direct effect in thediagnosis, cure, mitigation, treatment, or prevention of disease, or toaffect the structure or any function of the body of man or animals. Atherapeutic compound may be used in the form of a pharmaceuticallyacceptable salt, solvate, or solvate of a salt, e.g. the hydrochloride.Additionally, therapeutic compound may be provided as racemates, or asindividual enantiomers, including the R- or S-enantiomer. Thus, thetherapeutic compound may comprise a R-enantiomer only, a S-enantiomeronly, or a combination of both a R-enantiomer and a S-enantiomer of atherapeutic compound. In some aspects, the therapeutic compound may haveanti-inflammatory activity.

References herein to “therapeutic compound” may refer to isomyosmine oranother oxidoreductase inhibitor, an active compound other than aoxidoreductase inhibitor, or both.

In some aspects, isomyosmine or a pharmaceutically acceptable saltthereof may be administered for treating parasitic infections,non-limiting examples of which include malaria, African trypanosomiasis,babesiosis, Chagas disease, Giardia infections, leishmaniasis,roundworm, tapeworm, and toxoplasmosis. Many blood-borne parasites arespread by insects (vectors), so they are also referred to asvector-borne diseases. Malaria is a highly inflammatory disease causedby Plasmodium infection of host erythrocytes, an enzyme upregulatedduring malaria, which induce a strong inflammatory cytokine response inprimary human monocyte-derived macrophages. Elevated plasma xanthineoxidase (XO) activity correlates with high levels of inflammatorycytokines and with the development of cerebral malaria. See Ty et al.,“Malaria inflammation by xanthine oxidase-produced reactive oxygenspecies,” EMBO Mol Medv. 11 (8) (2019), PMC6685105.

In other aspects, isomyosmine or a pharmaceutically acceptable saltthereof may be administered for treating bacterial infections orbacterial diseases. Harmful bacteria that cause bacterial infections anddisease are called pathogenic bacteria. Bacterial diseases occur whenpathogenic bacteria get into the body and begin to reproduce and crowdout healthy bacteria, or to grow in tissues that are normally sterile.Harmful bacteria may also emit toxins that damage the body. Commonpathogenic bacteria and non-limiting examples of the types of bacterialdiseases they cause include Escherichia coli and Salmonella (foodpoisoning); Helicobacter pylori (gastritis and ulcers); Neisseriagonorrhoeae (gonorrhea); Neisseria meningitidis (meningitis);Staphylococcus aureus (boils, cellulitis, abscesses, wound infections,toxic shock syndrome, pneumonia, food poisoning); and Streptococcalbacteria (pneumonia, meningitis, ear infections, strep throat).Bacterial diseases are contagious and can result in many serious orlife-threatening complications, such as blood poisoning (bacteremia),kidney failure, and toxic shock syndrome.

In some examples, isomyosmine or a pharmaceutically acceptable saltthereof may be administered for treating Lyme disease. Lyme disease iscaused by the bacterium Borrelia burgdorferi and rarely, Borreliamayonii. If left untreated, infection can spread to joints, the heart,and the nervous system. Although Lyme disease may affect many organs,such as the heart and nervous system, joint involvement tends to be themost common and persistent manifestation, resulting in joint swellingand pain.

In other examples, isomyosmine or a pharmaceutically acceptable saltthereof may be administered for treating any indication associated withmortality or morbidity as identified in the Classification of Diseases(ICD) of the World Health Organization,https://www.who.int/clasSifications/icd/en/, the disclosure of which ishereby incorporated by reference in its entirety. Broadly, theseindications include infectious or parasitic diseases; neoplasms;diseases of the blood or blood-forming organs; diseases of the immunesystem; endocrine; nutritional or metabolic diseases; mental, behavioralor neurodevelopmental disorders; sleep-wake disorders; diseases of thenervous system; diseases of the visual system; diseases of the ear ormastoid process; diseases of the circulatory system; diseases of therespiratory system; diseases of the digestive system; diseases of theskin; diseases of the musculoskeletal system or connective tissue;diseases of the genitourinary system; conditions related to sexualhealth; pregnancy, childbirth or the puerperium; certain conditionsoriginating in the perinatal period; developmental anomalies; symptoms,signs or clinical findings, not elsewhere classified; injury, poisoningor certain other consequences of external causes. Non-limiting examplesof specific indications which may be treated are listed in the Diseases& Conditions A-Z Index, published by the Centers for Disease Control andPrevention (CDC), available athttps://www.cdc.gov/diseasesconditions/az/a.html, the disclosure ofwhich is hereby incorporated by reference in its entirety.

In some aspects, a therapeutic compound has an anti-inflammatoryactivity capable of reducing the levels of an inflammation-inducingmolecule. In some aspects, a therapeutic compound has ananti-inflammatory activity capable of reducing the levels of substanceP(SP), calcitonin gene-related peptide (CGRP), glutamate, or acombination thereof. In other aspects, a therapeutic compound has ananti-inflammatory activity capable of reducing the levels of SP, CGRP,glutamate, or a combination thereof released from a sensory neuron by,e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least30%, at least 35%, at least 40%, at least 45%, at least 50%, at least55%, at least 60%, at least 65%, at least 70%, at least 75%, at least80%, at least 85%, at least 90% or at least 95%. In yet other aspects, atherapeutic compound has an anti-inflammatory activity capable ofreducing the levels of SP, CGRP, glutamate, or a combination thereofreleased from a sensory neuron in a range from, e.g., about 10% to about100%, about 20% to about 100%, about 30% to about 100%, about 40% toabout 100%, about 50% to about 100%, about 60% to about 100%, about 70%to about 100%, about 80% to about 100%, about 10% to about 90%, about20% to about 90%, about 30% to about 90%, about 40% to about 90%, about50% to about 90%, about 60% to about 90%, about 70% to about 90%, about10% to about 80%, about 20% to about 80%, about 30% to about 80%, about40% to about 80%, about 50% to about 80%, or about 60% to about 80%,about 10% to about 70%, about 20% to about 70%, about 30% to about 70%,about 40% to about 70%, or about 50% to about 70%.

Prostaglandins mediate a local inflammatory response and are involved inall inflammatory functions through action on prostaglandin receptors andmediate inflammatory signaling including chemotaxis (macrophages,neutrophils and eosinophils), vasodilation and algesia. However, thePG-mediated inflammatory response is self-limiting (resolving). Theprinciple resolution factor is a prostaglandin called 15dPGJ2, which isan endogenous agonist of peroxisome proliferator-activator receptorgamma (PPAR-γ) signaling. PPAR-γ signaling pathway 1) induces apoptosisof macrophage M1 cells, thereby reducing the levels of Th1pro-inflammatory cytokines and 2) promotes differentiation of monocytesinto macrophage M2 cells. Macrophage M2 cells produce and release Th2anti-inflammatory cytokines.

In some aspects, a therapeutic compound has an anti-inflammatoryactivity capable of reducing the levels of an inflammation inducingprostaglandin. In other aspects, a therapeutic compound has ananti-inflammatory activity capable of reducing the levels of aninflammation inducing prostaglandin released from a sensory neuron by,e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least30%, at least 35%, at least 40%, at least 45%, at least 50%, at least55%, at least 60%, at least 65%, at least 70%, at least 75%, at least80%, at least 85%, at least 90% or at least 95%. In yet other aspects, atherapeutic compound has an anti-inflammatory activity capable ofreducing the levels of an inflammation inducing prostaglandin releasedfrom a sensory neuron in a range from, e.g., about 10% to about 100%,about 20% to about 100%, about 30% to about 100%, about 40% to about100%, about 50% to about 100%, about 60% to about 100%, about 70% toabout 100%, about 80% to about 100%, about 10% to about 90%, about 20%to about 90%, about 30% to about 90%, about 40% to about 90%, about 50%to about 90%, about 60% to about 90%, about 70% to about 90%, about 10%to about 80%, about 20% to about 80%, about 30% to about 80%, about 40%to about 80%, about 50% to about 80%, or about 60% to about 80%, about10% to about 70%, about 20% to about 70%, about 30% to about 70%, about40% to about 70%, or about 50% to about 70%.

In some aspects, a therapeutic compound has an anti-inflammatoryactivity substantially similar to 15dPGJ2. In some aspects, atherapeutic compound has an anti-inflammatory activity that is, e.g., atleast 5%, at least 15%, at least 25%, at least 50%, at least 55%, atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90% or at least 95% of the activity observed for15dPGJ2. In other aspects, a therapeutic compound has ananti-inflammatory activity that is in a range from, e.g., about 5% toabout 100%, about 50% to about 100%, about 60% to about 100%, about 70%to about 100%, about 80% to about 100%, about 25% to about 90%, about50% to about 90%, about 60% to about 90%, about 70% to about 90%, about80% to about 90%, about 25% to about 80%, about 50% to about 80%, about60% to about 80%, about 70% to about 80%, about 25% to about 70%, about50% to about 70%, about 25% to about 60%, about 50% to about 60%, orabout 25% to about 50% of the activity observed for 15dPGJ2.

The peroxisome proliferator-activated receptors (PPARs) are a group ofnuclear receptor proteins that function as transcription factorsregulating the expression of genes. All PPARs are known toheterodimerize with the retinoid X receptor (RXR) and bind to specificregions on the DNA of target genes called peroxisome proliferatorhormone response elements (PPREs). PPARs play essential roles in theregulation of cellular differentiation, development, and metabolism(carbohydrate, lipid, protein), and tumorigenesis of higher organisms.The family comprises three members, PPAR-α, PPAR-γ, and PPAR-δ (alsoknown as PPAR-β). PPAR-α is expressed in liver, kidney, heart, muscle,adipose tissue, as well as other tissues. PPAR-δ is expressed in manytissues but markedly in brain, adipose tissue, and skin. PPAR-γcomprises three alternatively-spliced forms, each with a differentexpression pattern. PPAR-γ1 is expressed in virtually all tissues,including heart, muscle, colon, kidney, pancreas, and spleen. PPAR-γ2 isexpressed mainly in adipose tissue. PPAR-γ3 is expressed in macrophages,large intestine, and white adipose tissue. Endogenous ligands for thePPARs include free fatty acids and eicosanoids. PPAR-γ is activated byPGJ2 (a prostaglandin), whereas PPAR-α is activated by leukotriene B4.

In some aspects, a therapeutic compound may have an anti-inflammatoryactivity capable of stimulating some or all PPAR signaling pathways. Itis contemplated that such a therapeutic compound therefore may act as aPPAR pan-agonist or possibly as a selective PPAR agonist.

In other aspects, a therapeutic compound has an anti-inflammatoryactivity capable of stimulating a PPAR-α signaling pathway. In someaspects, a therapeutic compound stimulates a PPAR-α signaling pathwayby, e.g., at least 5%, at least 15%, at least 25%, at least 50%, atleast 60%, at least 70%, at least 80%, or at least 90%. In otheraspects, a therapeutic compound stimulates a PPAR-α signaling pathway ina range from, e.g., about 5% to about 100%, about 50% to about 100%,about 60% to about 100%, about 70% to about 100%, about 80% to about100%, about 25% to about 90%, about 50% to about 90%, about 60% to about90%, about 70% to about 90%, about 80% to about 90%, about 25% to about80%, about 50% to about 80%, about 60% to about 80%, about 70% to about80%, about 25% to about 70%, about 50% to about 70%, about 25% to about60%, about 50% to about 60%, or about 25% to about 50%.

In some aspects, a therapeutic compound has an anti-inflammatoryactivity capable of stimulating a PPAR-6 signaling pathway. Atherapeutic compound may, for example, stimulate a PPAR-6 signalingpathway by at least 5%, at least 15%, at least 25%, at least 50%, atleast 60%, at least 70%, at least 80%, or at least 90%. In some cases, atherapeutic compound stimulates a PPAR-6 signaling pathway in a rangefrom, e.g., about 5% to about 100%, about 50% to about 100%, about 60%to about 100%, about 70% to about 100%, about 80% to about 100%, about25% to about 90%, about 50% to about 90%, about 60% to about 90%, about70% to about 90%, about 80% to about 90%, about 25% to about 80%, about50% to about 80%, about 60% to about 80%, about 70% to about 80%, about25% to about 70%, about 50% to about 70%, about 25% to about 60%, about50% to about 60%, or about 25% to about 50%.

In some aspects, a therapeutic compound has an anti-inflammatoryactivity capable of stimulating a PPAR-γ signaling pathway. Atherapeutic compound may be capable of binding to all isoforms ofPPAR-γ, or may be capable of selectively binding to either PPAR-γ1,PPAR-γ2, PPAR-γ3, or any combination of two thereof. A therapeuticcompound may stimulate a PPAR-γ signaling pathway by, e.g., at least 5%,at least 15%, at least 25%, at least 50%, at least 60%, at least 70%, atleast 80%, or at least 90%. A therapeutic compound may stimulate aPPAR-γ signaling pathway in a range from, e.g., about 5% to about 100%,about 50% to about 100%, about 60% to about 100%, about 70% to about100%, about 80% to about 100%, about 25% to about 90%, about 50% toabout 90%, about 60% to about 90%, about 70% to about 90%, about 80% toabout 90%, about 25% to about 80%, about 50% to about 80%, about 60% toabout 80%, about 70% to about 80%, about 25% to about 70%, about 50% toabout 70%, about 25% to about 60%, about 50% to about 60%, or about 25%to about 50%.

Macrophages are activated and polarized into distinct phenotypesexpressing unique cell surface molecules and secreting discrete sets ofcytokines and chemokines. The classical M1 phenotype supportspro-inflammatory Th1 responses driven by cytokines such as, e.g.,Interleukin-6 (IL-6), IL-12 and IL-23, while the alternate M2 phenotypeis generally supportive of anti-inflammatory processes driven by IL-10.M2 cells can be further classified into subsets, M2a, M2b, and M2c,based on the type of stimulation and the subsequent expression ofsurface molecules and cytokines.

In some aspects, a therapeutic compound has an anti-inflammatoryactivity capable of promoting the resolving phenotypic change of M1 toM2. A therapeutic compound may have an anti-inflammatory activitycapable of inducing apoptosis of macrophage M1 cells. A therapeuticcompound may have an anti-inflammatory activity capable of promotingdifferentiation of macrophage M2 cells. In yet another aspect, atherapeutic compound may have an anti-inflammatory activity capable ofinducing apoptosis of macrophage M1 cells and promoting differentiationof macrophage M2 cells.

In some aspects, a therapeutic compound has an anti-inflammatoryactivity capable of modulating Th1 and Th2 cytokines. A therapeuticcompound may have an anti-inflammatory activity capable of reducing thelevels of Interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α),interleukin-12 (IL-12), or a combination thereof released from a Th1cell. In other aspects, a therapeutic compound may have ananti-inflammatory activity capable of reducing the levels of IFN-γ,TNF-α, IL-12, or a combination thereof released from a Th1 cell by,e.g., at least 10%, at least 20%, at least 30%, at least 40%, at least50%, at least 60%, at least 70%, at least 80%, or at least 90%. In yetother aspects, a therapeutic compound may have an anti-inflammatoryactivity capable of reducing the levels of IFN-γ, TNF-α, IL-12, or acombination thereof released from a Th1 cell in a range from, e.g.,about 5% to about 100%, about 10% to about 100%, about 20% to about100%, about 30% to about 100%, about 40% to about 100%, about 50% toabout 100%, about 60% to about 100%, about 70% to about 100%, about 80%to about 100%, about 10% to about 90%, about 20% to about 90%, about 30%to about 90%, about 40% to about 90%, about 50% to about 90%, about 60%to about 90%, about 70% to about 90%, about 10% to about 80%, about 20%to about 80%, about 30% to about 80%, about 40% to about 80%, about 50%to about 80%, or about 60% to about 80%, about 10% to about 70%, about20% to about 70%, about 30% to about 70%, about 40% to about 70%, orabout 50% to about 70%.

In another aspect, a therapeutic compound has an anti-inflammatoryactivity capable of increasing the levels of IL-10 released from a Th2cell. A therapeutic compound may have an anti-inflammatory activitycapable of increasing the levels of IL-10 released from a Th2 cell by,e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least30%, at least 35%, at least 40%, at least 45%, at least 50%, at least55%, at least 60%, at least 65%, at least 70%, at least 75%, at least80%, at least 85%, at least 90% or at least 95%. In yet other aspects, atherapeutic compound may have an anti-inflammatory activity capable ofincreasing the levels of IL-10 released from a Th2 cell in a range from,e.g., about 5% to about 100%, about 10% to about 100%, about 20% toabout 100%, about 30% to about 100%, about 40% to about 100%, about 50%to about 100%, about 60% to about 100%, about 70% to about 100%, about80% to about 100%, about 10% to about 90%, about 20% to about 90%, about30% to about 90%, about 40% to about 90%, about 50% to about 90%, about60% to about 90%, about 70% to about 90%, about 10% to about 80%, about20% to about 80%, about 30% to about 80%, about 40% to about 80%, about50% to about 80%, or about 60% to about 80%, about 10% to about 70%,about 20% to about 70%, about 30% to about 70%, about 40% to about 70%,or about 50% to about 70%.

In another aspect, a therapeutic compound has an anti-inflammatoryactivity capable of reducing the levels of IFN-γ, TNF-α, IL-12, or acombination thereof released from a Th1 cell and increasing the levelsof IL-10 released from a Th2 cell. A therapeutic compound may have ananti-inflammatory activity capable of reducing the levels of IFN-γ,TNF-α, IL-12, or a combination thereof released from a Th1 cell by,e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least30%, at least 35%, at least 40%, at least 45%, at least 50%, at least55%, at least 60%, at least 65%, at least 70%, at least 75%, at least80%, at least 85%, at least 90% or at least 95%, and capable ofincreasing the levels of IL-10 released from a Th2 cell by, e.g., atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 55%, atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90% or at least 95%. In yet other aspects, atherapeutic compound may have an anti-inflammatory activity capable ofreducing the levels of IFN-γ, TNF-α, IL-12, or a combination thereofreleased from a Th1 cell in a range from, e.g., about 5% to about 100%,about 10% to about 100%, about 20% to about 100%, about 30% to about100%, about 40% to about 100%, about 50% to about 100%, about 60% toabout 100%, about 70% to about 100%, about 80% to about 100%, about 10%to about 90%, about 20% to about 90%, about 30% to about 90%, about 40%to about 90%, about 50% to about 90%, about 60% to about 90%, about 70%to about 90%, about 10% to about 80%, about 20% to about 80%, about 30%to about 80%, about 40% to about 80%, about 50% to about 80%, or about60% to about 80%, about 10% to about 70%, about 20% to about 70%, about30% to about 70%, about 40% to about 70%, or about 50% to about 70%, andcapable of increasing the levels of IL-10 released from a Th2 cell in arange from, e.g., about 10% to about 100%, about 20% to about 100%,about 30% to about 100%, about 40% to about 100%, about 50% to about100%, about 60% to about 100%, about 70% to about 100%, about 80% toabout 100%, about 10% to about 90%, about 20% to about 90%, about 30% toabout 90%, about 40% to about 90%, about 50% to about 90%, about 60% toabout 90%, about 70% to about 90%, about 10% to about 80%, about 20% toabout 80%, about 30% to about 80%, about 40% to about 80%, about 50% toabout 80%, or about 60% to about 80%, about 10% to about 70%, about 20%to about 70%, about 30% to about 70%, about 40% to about 70%, or about50% to about 70%.

In addition to oxidoreductase inhibitor(s), pharmaceutical formulationsmay include additional therapeutic compound(s) such as a non-steroidalanti-inflammatory drug (NSAID). NSAIDs are a large group of therapeuticcompounds with analgesic, anti-inflammatory, and anti-pyreticproperties. NSAIDs reduce inflammation by blocking cyclooxygenase.NSAIDs include, without limitation, aceclofenac, acemetacin, actarit,alcofenac, alminoprofen, amfenac, aloxipirin, aminophenazone,antraphenine, aspirin, azapropazone, benorilate, benoxaprofen,benzydamine, butibufen, celecoxib, chlorthenoxacin, choline salicylate,clometacin, dexketoprofen, diclofenac, diflunisal, emorfazone,epirizole; etodolac, etoricoxib, feclobuzone, felbinac, fenbufen,fenclofenac, flurbiprofen, glafenine, hydroxylethyl salicylate,ibuprofen, indometacin, indoprofen, ketoprofen, ketorolac, lactylphenetidin, loxoprofen, lumiracoxib, mefenamic acid, meloxicam,metamizole, metiazinic acid, mofebutazone, mofezolac, nabumetone,naproxen, nifenazone, niflumic acid, oxametacin, phenacetin, pipebuzone,pranoprofen, propyphenazone, proquazone, protizinic acid, rofecoxib,salicylamide, salsalate, sulindac, suprofen, tiaramide, tinoridine,tolfenamic acid, valdecoxib, and zomepirac.

NSAIDs may be classified based on their chemical structure or mechanismof action. Non-limiting examples of NSAIDs include a salicylatederivative NSAID, a p-amino phenol derivative NSAID, a propionic acidderivative NSAID, an acetic acid derivative NSAID, an enolic acidderivative NSAID, a fenamic acid derivative NSAID, a non-selectivecyclooxygenase (COX) inhibitor, a selective cyclooxygenase-1 (COX-1)inhibitor, and a selective cyclooxygenase-2 (COX-2) inhibitor. An NSAIDmay be a profen. Examples of a suitable salicylate derivative NSAIDinclude, without limitation, acetylsalicylic acid (aspirin), diflunisal,and salsalate. Examples of a suitable p-amino phenol derivative NSAIDinclude, without limitation, paracetamol and phenacetin. Examples of asuitable propionic acid derivative NSAID include, without limitation,alminoprofen, benoxaprofen, dexketoprofen, fenoprofen, flurbiprofen,ibuprofen, indoprofen, ketoprofen, loxoprofen, naproxen, oxaprozin,pranoprofen, and suprofen. Examples of a suitable acetic acid derivativeNSAID include, without limitation, aceclofenac, acemetacin, actarit,alcofenac, amfenac, clometacin, diclofenac, etodolac, felbinac,fenclofenac, indometacin, ketorolac, metiazinic acid, mofezolac,nabumetone, naproxen, oxametacin, sulindac, and zomepirac. Examples of asuitable enolic acid (oxicam) derivative NSAID include, withoutlimitation, droxicam, isoxicam, lornoxicam, meloxicam, piroxicam, andtenoxicam. Examples of a suitable fenamic acid derivative NSAID include,without limitation, flufenamic acid, mefenamic acid, meclofenamic acid,and tolfenamic acid. Examples of a suitable selective COX-2 inhibitorsinclude, without limitation, celecoxib, etoricoxib, firocoxib,lumiracoxib, meloxicam, parecoxib, rofecoxib, and valdecoxib.

A therapeutic compound may have a log P value indicating that thecompound is soluble in an organic solvent. As used herein, the term “logvalue” refers to the logarithm (base 10) of the partition coefficient(P) for a compound and is a measure of lipophilicity. Typically, P isdefined as the ratio of concentrations of a unionized compound in thetwo phases of a mixture of two immiscible solvents at equilibrium. Thus,log P=Log 10 (P), where P=[solute in immiscible solvent 1]/[solute inimmiscible solvent 2]. With regard to organic and aqueous phases, thelog P value of a compound is constant for any given pair of aqueous andorganic solvents, and its value can be determined empirically by one ofseveral phase-partitioning methods known to one skilled in the artincluding, e.g., a shake flask assay, a HPLC assay, and an interfacebetween two immiscible electrolyte solutions (ITIES) assay.

In some aspects, a therapeutic compound may have a log P valueindicating that the compound is substantially soluble in an organicsolvent. In some aspects, a therapeutic compound may have a log P valueindicating that the compound is, e.g., at least 50% soluble in anorganic solvent, at least 60% soluble in an organic solvent, at least70% soluble in an organic solvent, at least 80% soluble in an organicsolvent, or at least 90% soluble in an organic solvent. In some aspects,a therapeutic compound may have a log P value indicating that thecompound is between, e.g., about 50% to about 100% soluble in an organicsolvent, about 60% to about 100% soluble in an organic solvent, about70% to about 100% soluble in an organic solvent, about 80% to about 100%soluble in an organic solvent, or about 90% to about 100% soluble in anorganic solvent.

In some aspects, a therapeutic compound may have a log P value of, e.g.,more than 1.1, more than 1.2, more than 1.4, more than 1.6, more than1.8, more than 2.0, more than 2.2, more than 2.4, more than 2.6, morethan 2.8, more than 3.0, more than 3.2, more than 3.4, or more than 3.6.In other aspects of this embodiment, a therapeutic compound may have alog P value in the range of, e.g., between 1.8 and 4.0, between 2.0 and4.0, between 2.1 and 4.0, between 2.2 and 4.0, or between 2.3 and 4.0,between 2.4 and 4.0, between 2.5 and 4.0, between 2.6 and 4.0, orbetween 2.8 and 4.0. In other aspects of this embodiment, a therapeuticcompound may have a log P value in the range of, e.g., between 3.0 and4.0, or between 3.1 and 4.0, between 3.2 and 4.0, between 3.3 and 4.0,between 3.4 and 4.0, between 3.5 and 4.0, or between 3.6 and 4.0. Instill other aspects, a therapeutic compound may have a log P value inthe range of, e.g., between 2.0 and 2.5, between 2.0 and 2.7, between2.0 and 3.0, or between 2.2 and 2.5.

A therapeutic compound may have a polar surface area that ishydrophobic. As used herein, the term “polar surface area” refers to thesurface sum over all of the polar atoms in the structure of a compoundand is a measure of hydrophobicity. Typically, these polar atomsinclude, e.g., oxygen, nitrogen, and their attached hydrogens. In someaspects, a therapeutic compound may have a polar surface area of, e.g.,less than 8.0 nm², less than 7.0 nm², less than 6.0 nm², less than 5.0nm², less than 4.0 nm², or less than 3.0 nm².

In some aspects, a therapeutic compound may be a PPAR-γ agonist.Examples of a suitable PPAR-γ agonist include, without limitation,benzbromarone, a cannabidiol, cilostazol, curcumin,delta(9)-tetrahydrocannabinol, glycyrrhetinic acid, indomethacin,irbesartan, monascin, mycophenolic acid, resveratrol, 6-shogaol,telmisartan, a thiazolidinedione like rosiglitazone, pioglitazone, andtroglitazone, a NSAID, and a fibrate. Other suitable PPAR-γ agonists aredescribed in Masson et al. U.S. Pat. No. 8,461,183, the disclosure ofwhich is hereby incorporated by reference.

A therapeutic compound may be a nuclear receptor binding agent. Examplesof a suitable nuclear receptor binding agent include, withoutlimitation, a retinoic acid receptor (RAR) binding agent, a retinoid Xreceptor (RXR) binding agent, a liver X receptor (LXR) binding agent anda vitamin D binding agent.

A therapeutic compound may be an anti-hyperlipidemic agent. There areseveral classes of anti-hyperlipidemic agents (also known ashypolipidemic agents). They may differ in both their impact on thecholesterol profile and adverse effects. For example, some may lower lowdensity lipoprotein (LDL), while others may preferentially increase highdensity lipoprotein (HDL). Clinically, the choice of an agent willdepend on the cholesterol profile of an individual, cardiovascular riskof an individual, and/or the liver and kidney functions of anindividual. Examples of a suitable anti-hyperlipidemic agent include,without limitation, a fibrate, a statin, a tocotrienol, a niacin, a bileacid sequestrants (resin), a cholesterol absorption inhibitor, apancreatic lipase inhibitor, and a sympathomimetic amine.

A therapeutic compound may be a fibrate. Fibrates are a class ofamphipathic carboxylic acids with lipid level modifying properties.These therapeutic compounds are used for a range of metabolic disorders.One non-limiting use is as an anti-hyperlipidemic agent where it maylower levels of, e.g., triglycerides and LDL as well as increase levelsof HDL. Examples of a suitable fibrate include, without limitation,bezafibrate, ciprofibrate, clofibrate, gemfibrozil, and fenofibrate.

A therapeutic compound may be a statin. Statins (or HMG-CoA reductaseinhibitors) are a class of therapeutic compounds used to lower LDLand/or cholesterol levels by inhibiting the enzyme HMG-CoA reductase,which plays a central role in the production of cholesterol in theliver. To compensate for the decreased cholesterol availability,synthesis of hepatic LDL receptors is increased, resulting in anincreased clearance of LDL particles from the blood. Examples of asuitable statin include, without limitation, atorvastatin, fluvastatin,lovastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin.

A therapeutic compound may be a tocotrienol. Tocotrienols are anotherclass of HMG-CoA reductase inhibitors and may be used to lower LDLand/or cholesterol levels by inducing hepatic LDL receptor up-regulationand/or decreasing plasma LDL levels. Examples of a suitable tocotrienolinclude, without limitation, a γ-tocotrienol and a δ-tocotrienol.

A therapeutic compound may be a niacin. Niacins are a class oftherapeutic compounds with lipid level modifying properties. Forexample, a niacin may lower LDL by selectively inhibiting hepaticdiacyglycerol acyltransferase 2, reduce triglyceride synthesis, and VLDLsecretion through a receptor HM74 and HM74A or GPR109A. Thesetherapeutic compounds are used for a range of metabolic disorders. Onenon-limiting use is as an anti-hyperlipidemic agent where it may inhibitthe breakdown of fats in adipose tissue. Because a niacin blocks thebreakdown of fats, it causes a decrease in free fatty acids in the bloodand, as a consequence, decreases the secretion of very-low-densitylipoproteins (VLDL) and cholesterol by the liver. By lowering VLDLlevels, a niacin may also increase the level of HDL in blood. Examplesof a niacin include, without limitation, acipimox, niacin, nicotinamide,and vitamin B3.

A therapeutic compound may be a bile acid sequestrant. Bile acidsequestrants (also known as resins) are a class of therapeutic compoundsused to bind certain components of bile in the gastrointestinal tract.They disrupt the enterohepatic circulation of bile acids by sequesteringthem and preventing their reabsorption from the gut. Bile acidsequestrants are particularly effective for lowering LDL and cholesterolby sequestering the cholesterol-containing bile acids released into theintestine and preventing their reabsorption from the intestine. Inaddition, a bile acid sequestrant may also raise HDL levels. Examples ofa suitable bile acid sequestrant include, without limitation,cholestyramine, colesevelam, and colestipol.

In some aspects, a therapeutic compound may be a cholesterol absorptioninhibitor. Cholesterol absorption inhibitors are a class of therapeuticcompounds that inhibits the absorption of cholesterol from theintestine. Decreased cholesterol absorption leads to an up-regulation ofLDL-receptors on the surface of cells and an increased LDL-cholesteroluptake into these cells, thus decreasing levels of LDL in the bloodplasma. Examples of a suitable cholesterol absorption inhibitor include,without limitation, ezetimibe, a phytosterol, a sterol and a stanol.

A therapeutic compound may be a fat absorption inhibitor. Fat absorptioninhibitors are a class of therapeutic compounds that inhibits theabsorption of fat from the intestine. Decreased fat absorption reducescaloric intake. In one aspect, a fat absorption inhibitor inhibitspancreatic lipase, an enzyme that breaks down triglycerides in theintestine. Examples of a suitable fat absorption inhibitor include,without limitation, orlistat.

A therapeutic compound may be a sympathomimetic amine. Sympathomimeticamines are a class of therapeutic compounds that mimic the effects oftransmitter substances of the sympathetic nervous system such ascatecholamines, epinephrine (adrenaline), norepinephrine(noradrenaline), and/or dopamine. A sympathomimetic amine may act as anα-adrenergic agonist, a β-adrenergic agonist, a dopaminergic agonist, amonoamine oxidase (MAO) inhibitor, and a COMT inhibitor. Suchtherapeutic compounds, among other things, are used to treat cardiacarrest, low blood pressure, or even delay premature labor. Examples of asuitable sympathomimetic amine include, without limitation, clenbuterol,salbutamol, ephedrine, pseudoephedrine, methamphetamine, amphetamine,phenylephrine, isoproterenol, dobutamine, methylphenidate,lisdexamfetamine, cathine, cathinone, methcathinone, cocaine,benzylpiperazine (BZP), methylenedioxypyrovalerone (MDPV),4-methylaminorex, pemoline, phenmetrazine, and propylhexedrine.

A therapeutic compound may be an ester of a therapeutic compound. Ingeneral, an ester of a therapeutic compound increases the log P valuerelative to the same therapeutic compound without the estermodification. An ester group may be attached to a therapeutic compoundby, e.g., a carboxylic acid or hydroxyl functional group present of thetherapeutic compound. An ester of a therapeutic compound may have anincreased hydrophobicity, and as such, may be dissolved in a reducedvolume of solvent disclosed herein. In some instances, an ester of atherapeutic compound may be combined directly with an adjuvant disclosedherein, thereby eliminating the need of a solvent. An ester of atherapeutic compound may enable the making of a pharmaceuticalcomposition disclosed herein, in situations where a non-esterified formof the same therapeutic compound is otherwise immiscible in a solventdisclosed herein. An ester of a therapeutic compound may still bedelivered in a manner that more effectively inhibits a pro-inflammatoryresponse as long as the compound is combined with an adjuvant disclosedherein. In one embodiment, a therapeutic compound may be reacted withethyl ester in order to form an ethyl ester of the therapeutic compound.

In some aspects, a pharmaceutical composition does not comprise apharmaceutically acceptable solvent as previously described. Forexample, a pharmaceutical composition may comprise a therapeuticcompound and a pharmaceutically acceptable adjuvant but without apharmaceutically acceptable solvent.

A pharmaceutical composition may comprise a therapeutic compound in anamount sufficient to allow customary administration to an individual. Insome aspects, a pharmaceutical composition may be, e.g., at least 5 mg,at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least30 mg, at least 35 mg, at least 40 mg, at least 45 mg, at least 50 mg,at least 55 mg, at least 60 mg, at least 65 mg, at least 70 mg, at least75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg,or at least 100 mg of a therapeutic compound. In other aspects, apharmaceutical composition may be, e.g., at least 5 mg, at least 10 mg,at least 20 mg, at least 25 mg, at least 50 mg, at least 75 mg, at least100 mg, at least 200 mg, at least 300 mg, at least 400 mg, at least 500mg, at least 600 mg, at least 700 mg, at least 800 mg, at least 900 mg,at least 1,000 mg, at least 1,100 mg, at least 1,200 mg, at least 1,300mg, at least 1,400 mg, or at least 1,500 mg of a therapeutic compound.In yet other aspects, a pharmaceutical composition may be in the rangeof, e.g., about 5 mg to about 100 mg, about 10 mg to about 100 mg, about50 mg to about 150 mg, about 100 mg to about 250 mg, about 150 mg toabout 350 mg, about 250 mg to about 500 mg, about 350 mg to about 600mg, about 500 mg to about 750 mg, about 600 mg to about 900 mg, about750 mg to about 1,000 mg, about 850 mg to about 1,200 mg, or about 1,000mg to about 1,500 mg. In still other aspects, a pharmaceuticalcomposition may be in the range of, e.g., about 10 mg to about 250 mg,about 10 mg to about 500 mg, about 10 mg to about 750 mg, about 10 mg toabout 1,000 mg, about 10 mg to about 1,500 mg, about 50 mg to about 250mg, about 50 mg to about 500 mg, about 50 mg to about 750 mg, about 50mg to about 1,000 mg, about 50 mg to about 1,500 mg, about 100 mg toabout 250 mg, about 100 mg to about 500 mg, about 100 mg to about 750mg, about 100 mg to about 1,000 mg, about 100 mg to about 1,500 mg,about 200 mg to about 500 mg, about 200 mg to about 750 mg, about 200 mgto about 1,000 mg, about 200 mg to about 1,500 mg, about 5 mg to about1,500 mg, about 5 mg to about 1,000 mg, or about 5 mg to about 250 mg.

Pharmaceutical compositions may include a pharmaceutically acceptablesolvent. A solvent is a liquid, solid, or gas that dissolves anothersolid, liquid, or gaseous (the solute), resulting in a solution.Solvents useful in the pharmaceutical compositions include, withoutlimitation, a pharmaceutically acceptable polar aprotic solvent, apharmaceutically acceptable polar protic solvent and a pharmaceuticallyacceptable non-polar solvent. A pharmaceutically acceptable polaraprotic solvent includes, without limitation, dichloromethane (DCM),tetrahydrofuran (THF), ethyl acetate, acetone, dimethylformamide (DMF),acetonitrile (MeCN), dimethyl sulfoxide (DMSO). A pharmaceuticallyacceptable polar protic solvent includes, without limitation, aceticacid, formic acid, ethanol, n-butanol, 1-butanol, 2-butanol, isobutanol,sec-butanol, tert-butanol, n-propanol, isopropanol, 1,2 propan-diol,methanol, glycerol, and water. A pharmaceutically acceptable non-polarsolvent includes, without limitation, pentane, cyclopentane, hexane,cyclohexane, benzene, toluene, 1,4-dioxane, chloroform,n-methyl-pyrrilidone (NMP), and diethyl ether.

A pharmaceutical composition may comprise a solvent in an amountsufficient to dissolve a therapeutic compound disclosed herein. In otheraspects of this embodiment, a pharmaceutical composition may comprise asolvent in an amount of, e.g., less than about 90% (v/v), less thanabout 80% (v/v), less than about 70% (v/v), less than about 65% (v/v),less than about 60% (v/v), less than about 55% (v/v), less than about50% (v/v), less than about 45% (v/v), less than about 40% (v/v), lessthan about 35% (v/v), less than about 30% (v/v), less than about 25%(v/v), less than about 20% (v/v), less than about 15% (v/v), less thanabout 10% (v/v), less than about 5% (v/v), or less than about 1% (v/v).In other aspects of this embodiment, a pharmaceutical composition maycomprise a solvent in an amount in a range of, e.g., about 1% (v/v) to90% (v/v), about 1% (v/v) to 70% (v/v), about 1% (v/v) to 60% (v/v),about 1% (v/v) to 50% (v/v), about 1% (v/v) to 40% (v/v), about 1% (v/v)to 30% (v/v), about 1% (v/v) to 20% (v/v), about 1% (v/v) to 10% (v/v),about 2% (v/v) to 50% (v/v), about 2% (v/v) to 40% (v/v), about 2% (v/v)to 30% (v/v), about 2% (v/v) to 20% (v/v), about 2% (v/v) to 10% (v/v),about 4% (v/v) to 50% (v/v), about 4% (v/v) to 40% (v/v), about 4% (v/v)to 30% (v/v), about 4% (v/v) to 20% (v/v), about 4% (v/v) to 10% (v/v),about 6% (v/v) to 50% (v/v), about 6% (v/v) to 40% (v/v), about 6% (v/v)to 30% (v/v), about 6% (v/v) to 20% (v/v), about 6% (v/v) to 10% (v/v),about 8% (v/v) to 50% (v/v), about 8% (v/v) to 40% (v/v), about 8% (v/v)to 30% (v/v), about 8% (v/v) to 20% (v/v), about 8% (v/v) to 15% (v/v),or about 8% (v/v) to 12% (v/v).

In one embodiment, a solvent may comprise a pharmaceutically acceptablealcohol. As used herein, the term “alcohol” refers to an organicmolecule comprising a hydroxyl functional group (—OH) bonded to a carbonatom, where the carbon atom is saturated. In aspects of this embodiment,the alcohol may be, e.g., a C₁₋₄ alcohol, a C₂₋₄ alcohol, a C₁₋₅alcohol, a C₁₋₇ alcohol, a C₁₋₁₀ alcohol, a C₁₋₁₅ alcohol, or a C₁₋₂₀alcohol. In other aspects of this embodiment, an alcohol may be, e.g., aprimary alcohol, a secondary alcohol, or a tertiary alcohol. In otheraspects of this embodiment, an alcohol may be, e.g., an acyclic alcohol,a monohydric alcohol, a polyhydric alcohol (also known as a polyol orsugar alcohol), an unsaturated aliphatic alcohol, an alicyclic alcohol,or a combination thereof. Examples of a monohydric alcohol include,without limitation, methanol, ethanol, propanol, butanol, pentanol, and1-hexadecanol. Examples of a polyhydric alcohol include, withoutlimitation, glycol, glycerol, arabitol, erythritol, xylitol, maltitol,sorbitol (gluctiol), mannitol, inositol, lactitol, galactitol (iditol),and isomalt. Examples of an unsaturated aliphatic alcohol include,without limitation, prop-2-ene-1-ol, 3,7-dimethylocta-2,6-dien-1-ol, andprop-2-in-1-ol. Examples of an alicyclic alcohol include, withoutlimitation, cyclohexane-1,2,3,4,5,6-hexyl and2-(2-propyl)-5-methyl-cyclohexane-1-ol.

In another embodiment, a solvent may comprise an ester ofpharmaceutically acceptable alcohol and an acid. Suitablepharmaceutically acceptable alcohols include the ones disclosed herein.Suitable acids include, without limitation, acetic acid, butaric acid,and formic acid. An ester of an alcohol and an acid include, withoutlimitation, methyl acetate, methyl buterate, methyl formate, ethylacetate, ethyl buterate, ethyl formate, propyl acetate, propyl buterate,propyl formate, butyl acetate, butyl buterate, butyl formate, isobutylacetate, isobutyl buterate, isobutyl formate, pentyl acetate, pentylbuterate, pentyl formate, and 1-hexadecyl acetate, 1-hexadecyl buterate,and 1-hexadecyl formate.

In some aspects, a solvent may comprise a pharmaceutically acceptablepolyethylene glycol (PEG) polymer. PEG polymers, also known aspolyethylene oxide (PEO) polymers or polyoxyethylene (POE) polymers, areprepared by polymerization of ethylene oxide and are commerciallyavailable over a wide range of molecular weights from 100 g/mol to10,000,000 g/mol. PEG polymers with a low molecular mass are liquids orlow-melting solids, whereas PEG polymers of a higher molecular mass aresolids. A PEG polymer include, without limitation, PEG 100, PEG 200, PEG300, PEG 400, PEG 500, PEG 600, PEG 700, PEG 800, PEG 900, PEG 1000, PEG1100, PEG 1200, PEG 1300, PEG 1400, PEG 1500, PEG 1600, PEG 1700, PEG1800, PEG 1900, PEG 2000, PEG 2100, PEG 2200, PEG 2300, PEG 2400, PEG2500, PEG 2600, PEG 2700, PEG 2800, PEG 2900, PEG 3000, PEG 3250, PEG3350, PEG 3500, PEG 3750, PEG 4000, PEG 4250, PEG 4500, PEG 4750, PEG5000, PEG 5500, PEG 6000, PEG 6500, PEG 7000, PEG 7500, PEG 8000, PEG8500, PEG 9000, PEG 9500, PEG 10,000, PEG 11,000, PEG 12,000, PEG13,000, PEG 14,000, PEG 15,000, PEG 16,000, PEG 17,000, PEG 18,000, PEG19,000, or PEG 20,000.

In some aspects, a solvent may comprise a pharmaceutically acceptableglyceride. Glycerides comprise a substituted glycerol, where one, two,or all three hydroxyl groups of the glycerol are each esterified using afatty acid to produce monoglycerides, diglycerides, and triglycerides,respectively. In these compounds, each hydroxyl groups of glycerol maybe esterified by different fatty acids. Additionally, glycerides may beacetylated to produce acetylated monoglycerides, acetylateddiglycerides, and acetylated triglycerides.

In some aspects, a solvent may comprise a pharmaceutically acceptablesolid solvent. Solid solvents may be useful in the manufacture of asolid dose formulation of a pharmaceutical composition disclosed herein.Typically, a solid solvent is melted in order to dissolve a therapeuticcompound. A pharmaceutically acceptable solid solvent includes, withoutlimitation, menthol and PEG polymers described above.

An adjuvant is a pharmacological agent that modifies the effect of otheragents, such as one or more therapeutic compounds disclosed herein. Inaddition, an adjuvant may be used as a solvent that dissolves atherapeutic compound disclosed herein, forming an adjuvant solution. Anadjuvant may facilitate delivery of a therapeutic compound in a mannerthat more effectively inhibits a pro-inflammatory response. In someaspects, an adjuvant facilitates the delivery of a therapeutic compoundinto macrophages.

A pharmaceutical composition may comprise a pharmaceutically acceptableadjuvant in an amount sufficient to mix with a solution or an emulsion.In other aspects of this embodiment, a pharmaceutical composition maycomprise an adjuvant in an amount of, e.g., at least 10% (v/v), at least20% (v/v), at least 30% (v/v), at least 35% (v/v), at least 40% (v/v),at least 45% (v/v), at least 50% (v/v), at least 55% (v/v), at least 60%(v/v), at least 65% (v/v), at least 70% (v/v), at least 75% (v/v), atleast 80% (v/v), at least 85% (v/v), at least 90% (v/v), at least 95%(v/v), or at least 99% (v/v). In other aspects of this embodiment, apharmaceutical composition may comprise an adjuvant in an amount in arange of, e.g., about 30% (v/v) to about 99% (v/v), about 35% (v/v) toabout 99% (v/v), about 40% (v/v) to about 99% (v/v), about 45% (v/v) toabout 99% (v/v), about 50% (v/v) to about 99% (v/v), about 30% (v/v) toabout 98% (v/v), about 35% (v/v) to about 98% (v/v), about 40% (v/v) toabout 98% (v/v), about 45% (v/v) to about 98% (v/v), about 50% (v/v) toabout 98% (v/v), about 30% (v/v) to about 95% (v/v), about 35% (v/v) toabout 95% (v/v), about 40% (v/v) to about 95% (v/v), about 45% (v/v) toabout 95% (v/v), or about 50% (v/v) to about 95% (v/v). In yet otheraspects of this embodiment, a pharmaceutical composition may comprise anadjuvant in an amount in a range of, e.g., about 70% (v/v) to about 97%(v/v), about 75% (v/v) to about 97% (v/v), about 80% (v/v) to about 97%(v/v), about 85% (v/v) to about 97% (v/v), about 88% (v/v) to about 97%(v/v), about 89% (v/v) to about 97% (v/v), about 90% (v/v) to about 97%(v/v), about 75% (v/v) to about 96% (v/v), about 80% (v/v) to about 96%(v/v), about 85% (v/v) to about 96% (v/v), about 88% (v/v) to about 96%(v/v), about 89% (v/v) to about 96% (v/v), about 90% (v/v) to about 96%(v/v), about 75% (v/v) to about 93% (v/v), about 80% (v/v) to about 93%(v/v), about 85% (v/v) to about 93% (v/v), about 88% (v/v) to about 93%(v/v), about 89% (v/v) to about 93% (v/v), or about 90% (v/v) to about93% (v/v).

In some aspects, an adjuvant may be a pharmaceutically acceptable lipid.A lipid may be broadly defined as a hydrophobic or amphiphilic smallmolecule. The amphiphilic nature of some lipids allows them to formstructures such as vesicles, liposomes, or membranes in an aqueousenvironment. Non-limiting examples, of lipids include fatty acids,glycerolipids (like monoglycerides, diglycerides, and triglycerides),phospholipids, sphingolipids, sterol lipids, prenol lipids,saccharolipids, and polyketides. A pharmaceutical composition maycomprise a lipid such as, e.g. an oil, an oil-based liquid, a fat, afatty acid, a wax, a fatty acid ester, a fatty acid salt, a fattyalcohol, a glyceride (mono-, di- or tri-glyceride), a phospholipids, aglycol ester, a sucrose ester, a glycerol oleate derivative, a mediumchain triglyceride, or a mixture thereof.

A lipid useful in the pharmaceutical compositions may be apharmaceutically acceptable fatty acid. A fatty acid comprises acarboxylic acid with a long unbranched hydrocarbon chain which may beeither saturated or unsaturated. Thus arrangement confers a fatty acidwith a polar, hydrophilic end, and a nonpolar, hydrophobic end that isinsoluble in water. Most naturally occurring fatty acids have ahydrocarbon chain of an even number of carbon atoms, typically between 4and 24 carbons, and may be attached to functional groups containingoxygen, halogens, nitrogen, and sulfur. Synthetic or non-natural fattyacids may have a hydrocarbon chain of any number of carbon atoms frombetween 3 and 40 carbons. Where a double bond exists, there is thepossibility of either a cis or a trans geometric isomerism, whichsignificantly affects the molecule's molecular configuration. Cis-doublebonds cause the fatty acid chain to bend, an effect that is morepronounced the more double bonds there are in a chain. Most naturallyoccurring fatty acids are of the cis configuration, although the transform does exist in some natural and partially hydrogenated fats andoils. Examples of fatty acids include, without limitation, capryllicacid (8:0), pelargonic acid (9:0), capric acid (10:0), undecylic acid(11:0), lauric acid (12:0), tridecylic acid (13:0), myristic acid(14:0), myristoleic acid (14:1), pentadecyclic acid (15:0), palmiticacid (16:0), palmitoleic acid (16:1), sapienic acid (16:1), margaricacid (17:0), stearic acid (18:0), oleic acid (18:1), elaidic acid(18:1), vaccenic acid (18:1), linoleic acid (18:2), linoelaidic acid(18:2), α-linolenic acid (18:3), γ-linolenic acid (18:3), stearidonicacid (18:4), nonadecylic acid (19:0), arachidic acid (20:0), eicosenoicacid (20:1), dihomo-γ-linolenic acid (20:3), mead acid (20:3),arachidonic acid (20:4), eicosapentaenoic acid (20:5), heneicosylic acid(21:0), behenic acid (22:0), erucic acid (22:1), docosahexaenoic acid(22:6), tricosylic acid (23:0), lignoceric acid (24:0), nervonic acid(24:1), pentacosylic acid (25:0), cerotic acid (26:0), heptacosylic acid(27:0), montanic acid (28:0), nonacosylic acid (29:0), melissic acid(30:0), henatriacontylic acid (31:0), lacceroic acid (32:0), psyllicacid (33:0), geddic acid (34:0), ceroplastic acid (35:0), andhexatriacontylic acid (36:0).

In an embodiment, an adjuvant may be a pharmaceutically acceptablesaturated or unsaturated fatty acid. A saturated or unsaturated fattyacid may comprise, e.g., at least 8, at least 10, at least 12, at least14, at least 16, at least 18, at least 20, at least 22, at least 24, atleast 26, at least 28, or at least 30 carbon atoms. In some instances, asaturated or unsaturated fatty acid comprises, e.g., between 4 and 24carbon atoms, between 6 and 24 carbon atoms, between 8 and 24 carbonatoms, between 10 and 24 carbon atoms, between 12 and 24 carbon atoms,between 14 and 24 carbon atoms, or between 16 and 24 carbon atoms,between 4 and 22 carbon atoms, between 6 and 22 carbon atoms, between 8and 22 carbon atoms, between 10 and 22 carbon atoms, between 12 and 22carbon atoms, between 14 and 22 carbon atoms, or between 16 and 22carbon atoms, between 4 and 20 carbon atoms, between 6 and 20 carbonatoms, between 8 and 20 carbon atoms, between 10 and 20 carbon atoms,between 12 and 20 carbon atoms, between 14 and 20 carbon atoms, orbetween 16 and 20 carbon atoms. If unsaturated, the fatty acid may have,e.g., 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, or 6 ormore double bonds.

A pharmaceutically acceptable saturated or unsaturated fatty acid may beliquid at room temperature. The melting point of a fatty acid is largelydetermined by the degree of saturation/unsaturation of the hydrocarbonchain. In aspects of this embodiment, a saturated or unsaturated fattyacid has a melting point temperature of, e.g., 20° C. or below, 15° C.or below, 10° C. or below, 5° C. or below, 0° C. or below, −5° C. orbelow, −10° C. or below, −15° C. or below, or −20° C. or below. In otheraspects of this embodiment, a saturated or unsaturated fatty acid has amelting point temperature in the range of, e.g., about −20° C. to about20° C., about −20° C. to about 18° C., about −20° C. to about 16° C.,about −20° C. to about 12° C., about −20° C. to about 8° C., about −20°C. to about 4° C., about −20° C. to about 0° C., about −15° C. to about20° C., about −15° C. to about 18° C., about −15° C. to about 16° C.,about −15° C. to about 12° C., about −15° C. to about 8° C., about −15°C. to about 4° C., or about −15° C. to about 0° C.

In some aspects, an adjuvant may comprise one kind of pharmaceuticallyacceptable fatty acid. An adjuvant may comprise, for example, onlypalmitic acid, only stearic acid, only oleic acid, only linoleic acid,or only linolenic acid. Alternatively, an adjuvant may comprise aplurality of different pharmaceutically acceptable fatty acids. Anadjuvant may comprise, e.g., two or more different fatty acids, three ormore different fatty acids, four or more different fatty acids, five ormore different fatty acids, or six or more different fatty acids.

In some aspects, an adjuvant may comprise two or more differentpharmaceutically acceptable fatty acids including at least palmiticacid, stearic acid, oleic acid, linoleic acid and/or linolenic acid, andany combination thereof. An adjuvant may comprise a ratio of palmiticacid and/or stearic acid and/or oleic acid:linolenic acid and/orlinoleic acid of, e.g., at least 2:1, at least 3:1, at least 4:1, atleast 5:1, at least 6:1, at least 7:1, at least 8:1, at least 9:1, atleast 10:1, at least 15:1, or at least 20:1. In some examples, anadjuvant may comprise a ratio of palmitic acid and/or stearic acidand/or oleic acid:linolenic acid and/or linoleic acid in a range of,e.g., about 1:1 to about 20:1, about 2:1 to about 15:1, about 4:1 toabout 12:1, or about 6:1 to about 10:1.

In other aspects, an adjuvant may comprise four or more differentpharmaceutically acceptable fatty acids including at least palmiticacid, stearic acid, oleic acid, linoleic acid and/or linolenic acid, andany combination thereof. In other aspects of this embodiment, anadjuvant may comprise a ratio of palmitic acid:stearic acid:linolenicacid:linoleic acid of, e.g., 10:10:1:1, 9:9:1:1, 8:8:1:1, 7:7:1:1,6:6:1:1, 5:5:1:1, 4:4:1:1, 3:3:1:1, 2:2:1:1, or 1:1:1:1. In otheraspects of this embodiment, an adjuvant may comprise a ratio of palmiticacid:stearic acid:linolenic acid:linoleic acid in a range of, e.g.,about 10:10:1:1 to about 6:6:1:1, about 8:8:1:1 to about 4:4:1:1, orabout 5:5:1:1 to about 1:1:1:1.

A lipid useful in the pharmaceutical compositions may be apharmaceutically acceptable omega fatty acid. Non-limiting examples ofan omega fatty acid include omega-3, omega-6, and omega-9. Omega-3 fattyacids (also known as n-3 fatty acids or ω-3 fatty acids) are a family ofessential unsaturated fatty acids that have in common a finalcarbon-carbon double bond in the n-3 position, that is, the third bond,counting from the methyl end of the fatty acid. The omega-3 fatty acidsare “essential” fatty acids because they are vital for normal metabolismand cannot be synthesized by the human body. An omega-3 fatty acidincludes, without limitation, hexadecatrienoic acid (16:3), α-linolenicacid (18:3), stearidonic acid (18:4), eicosatrienoic acid (20:3),eicosatetraenoic acid (20:4), eicosapentaenoic acid (20:5),heneicosapentaenoic acid (21:5), docosapentaenoic acid (22:5),clupanodonic acid (22:5), docosahexaenoic acid (22:6),tetracosapentaenoic acid (24:5), and tetracosahexaenoic acid (nisinicacid) (24:6).

Omega-6 fatty acids (also known as n-6 fatty acids or ω-6 fatty acids)are a family of unsaturated fatty acids that have in common a finalcarbon-carbon double bond in the n-6 position, that is, the sixth bond,counting from the methyl end of the fatty acid. An omega-6 fatty acidincludes, without limitation, linoleic acid (18:2), gamma-linolenic acid(18:3), calendic acid (18:3), eicosadienoic acid (20:2),dihomo-gamma-linolenic acid (20:3), arachidonic acid (20:4),docosadienoic acid (22:2), adrenic acid (22:4), docosapentaenoic acid(22:5), tetracosatetraenoic acid (24:4), and tetracosapentaenoic acid(24:5). Omega-9 fatty acids (also known as n-9 fatty acids or ω-9 fattyacids) are a family of unsaturated fatty acids that have in common afinal carbon-carbon double bond in the n-9 position, that is, the ninthbond, counting from the methyl end of the fatty acid. An omega-9 fattyacid includes, without limitation, oleic acid (18:1), elaidic acid(18:1), eicosenoic acid (20:1), mead acid (20:3), erucic acid (22:1),and nervonic acid (24:1).

A lipid useful in the pharmaceutical compositions may be apharmaceutically acceptable oil. An oil includes any fatty acid that isliquid at normal room temperature, such as, e.g. about 20° C. Incontrast, a fat includes any fatty acid that is solid at normal roomtemperature, such as, e.g. about 20° C. An oil suitable as a lipiduseful in the pharmaceutical compositions disclosed herein, may be anatural oil or a vegetable oil. Examples of suitable natural oilsinclude, without limitation, mineral oil, triacetin, ethyl oleate, ahydrogenated natural oil, or a mixture thereof. Examples of suitablevegetable oils include, without limitation, almond oil, arachis oil,avocado oil, canola oil, castor oil, coconut oil, corn oil, cottonseedoil, grape seed oil, hazelnut oil, hemp oil, linseed oil (flax seedoil), olive oil, palm oil, peanut oil, rapeseed oil, rice bran oil,safflower oil, sesame oil, soybean oil, soya oil, sunflower oil, walnutoil, wheat germ oil, or a mixture thereof. Each of these oils iscommercially available from a number of sources well recognized by thoseskilled in the art.

An oil is typically a mixture of various fatty acids. For example,rapeseed oil, obtained from the seeds of Brassica napus, includes bothomega-6 and omega-3 fatty acids in a ratio of about 2:1. As anotherexample, linseed oil, obtained from the seeds of Linum usitatissimum,includes about 7% palmitic acid, about 3.4-4.6% stearic acid, about18.5-22.6% oleic acid, about 14.2-17% linoleic acid, and about51.9-55.2% α-linolenic acid. In some instances, a pharmaceuticalcomposition comprises an oil including at least two different fattyacids, at least three different fatty acids, at least four differentfatty acids, at least five different fatty acids, or at least sixdifferent fatty acids.

A lipid useful in the pharmaceutical compositions may be apharmaceutically acceptable glycerolipid. Glycerolipids are composedmainly of mono-, di-, and tri-substituted glycerols. One group ofglycerolipids is the glycerides, where one, two, or all three hydroxylgroups of glycerol are each esterified using a fatty acid to producemonoglycerides, diglycerides, and triglycerides, respectively. In thesecompounds, each hydroxyl groups of glycerol may be esterified bydifferent fatty acids. Additionally, glycerides may be acetylated toproduce acetylated monoglycerides, acetylated diglycerides, andacetylated triglycerides. One group of glycerolipids is the glycerides,where one, two, or all three hydroxyl groups of glycerol have sugarresidues attached via a glycosidic linkage.

In some instances, compositions may include one or more pharmaceuticallyacceptable stabilizing agents. A stabilizing agent reduces or eliminatesformation of esters of a therapeutic compound that may result as aunwanted reaction with the particular solvent used. A stabilizing agentinclude, without limitation, water, a sacrificial acid comprising afatty acid component and acetic acid, ethyl acetate, a sodiumacetate/acetic acid (E262), a monoglyceride, an acetylatedmonoglyceride, a diglyceride, an acetylated monoglyceride, an acetylateddiglyceride, a fatty acid, and a fatty acid salt.

In some aspects, a pharmaceutically acceptable stabilizing agent maycomprise a pharmaceutically acceptable emulsifying agent. An emulsifyingagent (also known as an emulgent) is a substance that stabilizes anemulsion comprising a liquid dispersed phase and a liquid continuousphase by increasing its kinetic stability. Thus, in situations where thesolvent and adjuvant used to make a pharmaceutical composition arenormally immiscible, an emulsifying agent is used to create a homogenousand stable emulsion. An emulsifying agent includes, without limitation,a surfactant, a polysaccharide, a lectin, and a phospholipid.

In some aspects, an emulsifying agent may comprise a surfactant. As usedhereon, the term “surfactant” refers to a natural or syntheticamphiphilic compound. A surfactant can be non-ionic, zwitterionic, orionic. Non-limiting examples of surfactants include polysorbates likepolysorbate 20 (TWEEN® 20), polysorbate 40 (TWEEN® 40), polysorbate 60(TWEEN® 60), polysorbate 61 (TWEEN® 61), polysorbate 65 (TWEEN® 65),polysorbate 80 (TWEEN® 80), and polysorbate 81 (TWEEN® 81); poloxamers(polyethylene-polypropylene copolymers), such as Poloxamer 124(PLURONIC® L44), Poloxamer 181 (PLURONIC® L61), Poloxamer 182 (PLURONIC®L62), Poloxamer 184 (PLURONIC® L64), Poloxamer 188 (PLURONIC® F68),Poloxamer 237 (PLURONIC® F87), Poloxamer 338 (PLURONIC® L108), Poloxamer407 (PLURONIC® F127), polyoxyethyleneglycol dodecyl ethers, such asBRIJ® 30, and BRIJ® 35; 2-dodecoxyethanol (LUBROL®-PX); polyoxyethyleneoctyl phenyl ether (TRITON® X-100); sodium dodecyl sulfate (SDS);3[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS);3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulfonate(CHAPSO); sucrose monolaurate; and sodium cholate. Other non-limitingexamples of surfactant excipients can be found in, e.g., Ansel, supra,(1999); Gennaro, supra, (2000); Hardman, supra, (2001); and Rowe, supra,(2003), each of which is hereby incorporated by reference in itsentirety.

In some aspects, an emulsifying agent may comprise a polysaccharide.Non-limiting examples of polysaccharides include guar gum, agar,alginate, calgene, a dextran (like dextran 1K, dextran 4K, dextran 40K,dextran 60K, and dextran 70K), dextrin, glycogen, inulin, starch, astarch derivative (like hydroxymethyl starch, hydroxyethyl starch,hydroxypropyl starch, hydroxybutyl starch, and hydroxypentyl starch),hetastarch, cellulose, FICOLL, methyl cellulose (MC), carboxymethylcellulose (CMC), hydroxyethyl cellulose (HEC), hydroxypropyl cellulose(HPC), hydroxyethyl methyl cellulose (NEMC), hydroxypropyl methylcellulose (HPMC); polyvinyl acetates (PVA); polyvinyl pyrrolidones(PVP), also known as povidones, having a K-value of less than or equalto 18, a K-value greater than 18 or less than or equal to 95, or aK-value greater than 95, like PVP 12 (KOLLIDON® 12), PVP 17 (KOLLIDON®17), PVP 25 (KOLLIDON® 25), PVP 30 (KOLLIDON® 30), PVP 90 (KOLLIDON®90); and polyethylene imines (PEI).

In some aspects, an emulsifying agent may comprise a lectin. Lectins aresugar-binding proteins that are highly specific for their sugarmoieties. Lectins may be classified according to the sugar moiety thatthey bind to, and include, without limitation, mannose-binding lectins,galactose/N-acetylgalactosamine-binding lectins,N-acetylgluxosamine-binding lectins, N-acetylneuramine-binding lectins,N-acetylneuraminic acid-binding lectins, and fucose-binding lectins.Non-limiting examples of surfactants include concanavain A, lentillectin, snowdrop lectin, Roin, peanut agglutinin, jacain, hairy vetchlectin, wheat germ agglutinin, elderberry lectin, Maackia anurensisleukoagglutinin, Maackia anurensis hemoagglutinin, Ulex europaeusagglutinin, and Aleuria aurantia lectin.

In some aspects, an emulsifying agent may comprise a phospholipid. Thestructure of the phospholipid generally comprises a hydrophobic tail anda hydrophilic head and is amphipathic in nature. Most phospholipidscontain a diglyceride, a phosphate group, and a simple organic moleculesuch as choline; one exception to this rule is sphingomyelin, which isderived from sphingosine instead of glycerol. Phospholipids include,without limitation, diacylglycerides and phosphosphingolipids.Non-limiting examples of diacylglycerides include a phosphatidic acid(phosphatidate) (PA), a phosphatidylethanolamine (cephalin) (PE), aphosphatidylcholine (lecithin) (PC), a phosphatidylserine (PS), and aphosphoinositide including phosphatidylinositol (PI),phosphatidylinositol phosphate (PIP), phosphatidylinositol bisphosphate(PIP2), and phosphatidylinositol triphosphate (PIP3). Non-limitingexamples of phosphosphingolipids include a ceramide phosphorylcholine(sphingomyelin) (SPH), ceramide phosphorylethanolamine (sphingomyelin)(Cer-PE), and ceramide phosphorylglycerol.

The pharmaceutical compositions may act as a delivery system thatenables the therapeutic compound(s) to be more effectively delivered ortargeted to a cell type, tissue, organ, or region of the body in amanner that more effectively inhibits a pro-inflammatory response. Thisinhibition results in an improved treatment of a chronic inflammation.For example, a pharmaceutical composition may facilitate the delivery ofa therapeutic compound into macrophages. One possible mechanism thatachieves this selective biodistribution is that the pharmaceuticalcompositions may be designed to take advantage of the activity ofchylomicrons. Chylomicrons are relatively large lipoprotein particleshaving a diameter of 75 nm to 1,200 nm. Comprising triglycerides(85-92%), phospholipids (6-12%), cholesterol (1-3%) and apolipoproteins(1-2%), chylomicrons transport dietary lipids from the intestines toother locations in the body. Chylomicrons are one of the five majorgroups of lipoproteins, the others being VLDL, IDL, low-densitylipoproteins (LDL), high-density lipoproteins (HDL), that enable fatsand cholesterol to move within the water-based solution of thebloodstream.

During digestion, fatty acids and cholesterol undergo processing in thegastrointestinal tract by the action of pancreatic juices includinglipases and emulsification with bile salts to generate micelles. Thesemicelles allow the absorption of lipid as free fatty acids by theabsorptive cells of the small intestine, known as enterocytes. Once inthe enterocytes, triglycerides and cholesterol are assembled intonascent chylomicrons. Nascent chylomicrons are primarily composed oftriglycerides (85%) and contain some cholesterol and cholesteryl esters.The main apolipoprotein component is apolipoprotein B-48 (APOB48). Thesenascent chylomicrons are released by exocytosis from enterocytes intolacteals, lymphatic vessels originating in the villi of the smallintestine, and are then secreted into the bloodstream at the thoracicduct's connection with the left subclavian vein.

While circulating in lymph and blood, chylomicrons exchange componentswith HDL. The HDL donates apolipoprotein C-II (APOC2) and apolipoproteinE (APOE) to the nascent chylomicron and thus converts it to a maturechylomicron (often referred to simply as “chylomicron”). APOC2 is thecofactor for lipoprotein lipase (LPL) activity. Once triglyceride storesare distributed, the chylomicron returns APOC2 to the HDL (but keepsAPOE), and, thus, becomes a chylomicron remnant, now only 30-50 nm.APOB48 and APOE are important to identify the chylomicron remnant in theliver for endocytosis and breakdown into lipoproteins (VLDL, LDL andHDL). These lipoproteins are processed and stored by competent cells,including, e.g., hepatocytes, adipocytes and macrophages. Thus, withoutwishing to be limited by any theory, upon oral administration of thepharmaceutical compositions are processed into micelles while in thegastrointestinal tract, absorbed by enterocytes and assembled intonascent chylomicrons, remain associated with chylomicron remnants takenup by the liver, and ultimately loaded into macrophages.

In some aspects, a method of preparing a pharmaceutical composition isprovided. A method may comprise the step of contacting apharmaceutically acceptable adjuvant with a therapeutic compound underconditions which allow the therapeutic compound to dissolve in thepharmaceutically acceptable adjuvant, thereby forming a pharmaceuticalcomposition.

Other aspects include a method of preparing a pharmaceuticalcomposition. A method may comprise the steps of a) contacting apharmaceutically acceptable solvent with a therapeutic compound underconditions which allow the therapeutic compound to dissolve in thepharmaceutically acceptable solvent, thereby forming a solution; and b)contacting the solution formed in step (a) with a pharmaceuticallyacceptable adjuvant under conditions which allow the formation of apharmaceutical composition. The methods of preparing may furthercomprise a step (c) of removing the pharmaceutically acceptable solventfrom the pharmaceutical composition.

The amount of therapeutic compound that is contacted with thepharmaceutically acceptable solvent in step (a) of the method may varywidely. Factors that may influence the amount of a therapeutic compoundused include, among others, the final amount the therapeutic compounddesired in the pharmaceutical composition, the desired concentration ofa therapeutic compound in the solution, the hydrophobicity of thetherapeutic compound, the lipophobicity of the therapeutic compound, thetemperature under which the contacting step (a) is performed, and thetime under which the contacting step (a) is performed.

The volume of a pharmaceutically acceptable solvent used in step (a) ofthe method also may vary over a wide range. Factors that may influencethe volume of pharmaceutically acceptable solvent used include, amongothers, the final amount of pharmaceutical composition desired, thedesired concentration of a therapeutic compound in the solution, thehydrophobicity of the therapeutic compound, and the lipophobicity of thetherapeutic compound.

In aspects of this embodiment, the amount of a therapeutic compound thatis contacted with the solvent in step (a) may be, e.g., at least 10 mg,at least 20 mg, at least 30 mg, at least 40 mg, at least 50 mg, at least60 mg, at least 70 mg, at least 80 mg, at least 90 mg, at least 100 mg,at least 200 mg, at least 300 mg, at least 400 mg, at least 500 mg, atleast 600 mg, at least 700 mg, at least 800 mg, at least 900 mg, atleast 1,000 mg, at least 1,100 mg, at least 1,200 mg, at least 1,300 mg,at least 1,400 mg, or at least 1,500 mg. In other aspects of thisembodiment, the amount of a therapeutic compound that is contacted withthe solvent in step (a) may be in the range of, e.g., about 10 mg toabout 100 mg, about 50 mg to about 150 mg, about 100 mg to about 250 mg,about 150 mg to about 350 mg, about 250 mg to about 500 mg, about 350 mgto about 600 mg, about 500 mg to about 750 mg, about 600 mg to about 900mg, about 750 mg to about 1,000 mg, about 850 mg to about 1,200 mg, orabout 1,000 mg to about 1,500 mg. In other aspects of this embodiment,the amount of a therapeutic compound that is dissolved in the solvent instep (a) may be in the range of, e.g., about 10 mg to about 250 mg,about 10 mg to about 500 mg, about 10 mg to about 750 mg, about 10 mg toabout 1,000 mg, about 10 mg to about 1,500 mg, about 50 mg to about 250mg, about 50 mg to about 500 mg, about 50 mg to about 750 mg, about 50mg to about 1,000 mg, about 50 mg to about 1,500 mg, about 100 mg toabout 250 mg, about 100 mg to about 500 mg, about 100 mg to about 750mg, about 100 mg to about 1,000 mg, about 100 mg to about 1,500 mg,about 200 mg to about 500 mg, about 200 mg to about 750 mg, about 200 mgto about 1,000 mg, or about 200 mg to about 1,500 mg.

Step (a) may be carried out at room temperature, in order to allow atherapeutic compound to dissolve fully in the pharmaceuticallyacceptable solvent. However, in other embodiments of the method, step(a) may be carried out at a temperature that is greater than roomtemperature, e.g., greater than 21° C., greater than 25° C., greaterthan 30° C., greater than 35° C. or greater than 37° C. In certaincases, Step (a) may be carried out at temperatures below roomtemperature, in order to allow a therapeutic compound to dissolve fullyin solvent. However, in other embodiments of the method, step (a) may becarried out at a temperature that is less than room temperature, e.g.,less than 10° C., greater than 5° C., greater than 0° C., greater than−10° C. or greater than −20° C. The contacting in Step (a) may comprisemixing the therapeutic compound and the pharmaceutically acceptablesolvent, e.g., by stirring, inversion, sonication, or vortexing. Themixing may be carried out for, e.g., at least 1 second, at least 5seconds, at least 10 seconds, at least 20 seconds, at least 30 seconds,at least 45 seconds, at least 60 seconds, or more, until the therapeuticcompound is fully dissolved in the solvent.

The concentration of a therapeutic compound in a solution may vary overa wide range. By way of example, the concentration of the therapeuticcompound may be at least 0.00001 mg/mL, at least 0.0001 mg/mL, at least0.001 mg/mL, at least 0.01 mg/mL, at least 0.1 mg/mL, at least 1 mg/mL,at least 10 mg/mL, at least 25 mg/mL, at least 50 mg/mL, at least 100mg/mL, at least 200 mg/mL, at least 500 mg/mL, at least 700 mg/mL, atleast 1,000 mg/mL, or at least 1,200 mg/mL. The concentration of thetherapeutic compound may be, e.g., at most 1,000 mg/mL, at most 1,100mg/mL, at most 1,200 mg/mL, at most 1,300 mg/mL, at most 1,400 mg/mL, atmost 1,500 mg/mL, at most 2,000 mg/mL, at most 2,000 mg/mL, or at most3,000 mg/mL. In some instances, the concentration of a therapeuticcompound may be in a range of, e.g., about 0.00001 mg/mL to about 3,000mg/mL, about 0.0001 mg/mL to about 3,000 mg/mL, about 0.01 mg/mL toabout 3,000 mg/mL, about 0.1 mg/mL to about 3,000 mg/mL, about 1 mg/mLto about 3,000 mg/mL, about 250 mg/mL to about 3,000 mg/mL, about 500mg/mL to about 3,000 mg/mL, about 750 mg/mL to about 3,000 mg/mL, about1,000 mg/mL to about 3,000 mg/mL, about 100 mg/mL to about 2,000 mg/mL,about 250 mg/mL to about 2,000 mg/mL, about 500 mg/mL to about 2,000mg/mL, about 750 mg/mL to about 2,000 mg/mL, about 1,000 mg/mL to about2,000 mg/mL, about 100 mg/mL to about 1,500 mg/mL, about 250 mg/mL toabout 1,500 mg/mL, about 500 mg/mL to about 1,500 mg/mL, about 750 mg/mLto about 1,500 mg/mL, about 1,000 mg/mL to about 1,500 mg/mL, about 100mg/mL to about 1,200 mg/mL, about 250 mg/mL to about 1,200 mg/mL, about500 mg/mL to about 1,200 mg/mL, about 750 mg/mL to about 1,200 mg/mL,about 1,000 mg/mL to about 1,200 mg/mL, about 100 mg/mL to about 1,000mg/mL, about 250 mg/mL to about 1,000 mg/mL, about 500 mg/mL to about1,000 mg/mL, about 750 mg/mL to about 1,000 mg/mL, about 100 mg/mL toabout 750 mg/mL, about 250 mg/mL to about 750 mg/mL, about 500 mg/mL toabout 750 mg/mL, about 100 mg/mL to about 500 mg/mL, about 250 mg/mL toabout 500 mg/mL, about 0.00001 mg/mL to about 0.0001 mg/mL, about0.00001 mg/mL to about 0.001 mg/mL, about 0.00001 mg/mL to about 0.01mg/mL, about 0.00001 mg/mL to about 0.1 mg/mL, about 0.00001 mg/mL toabout 1 mg/mL, about 0.001 mg/mL to about 0.01 mg/mL, about 0.001 mg/mLto about 0.1 mg/mL, about 0.001 mg/mL to about 1 mg/mL, about 0.001mg/mL to about 10 mg/mL, or about 0.001 mg/mL to about 100 mg/mL.

The volume of a pharmaceutically acceptable adjuvant used in step (b) ofthe method may be any volume desired. Factors used to determine thevolume of a pharmaceutically acceptable adjuvant used include, withoutlimitation, the final amount of a pharmaceutical composition desired,the desired concentration of a therapeutic compound in thepharmaceutical composition, the ratio of solvent:adjuvant used, and themiscibility of solvent and adjuvant.

In aspects of this embodiment, the ratio of solution:adjuvant may be,e.g., at least 5:1, at least 4:1, at least 3:1, at least 2:1, at least0:1, at least 1:1, at least 1:2, at least 1:3, at least 1:4, at least1:5, at least 1:6, at least 1:7, at least 1:8, at least 1:9, at least1:10, at least 1:15, at least 1:20, or at least 1:25. In other aspectsof this embodiment, the ratio of solution:adjuvant may be in a range of,e.g., about 5:1 to about 1:25, about 4:1 to about 1:25, about 3:1 toabout 1:25, about 2:1 to about 1:25, about 0:1 to about 1:25, about 1:1to about 1:25, about 1:2 to about 1:25, about 1:3 to about 1:25, about1:4 to about 1:25, about 1:5 to about 1:25, about 5:1 to about 1:20,about 4:1 to about 1:20, about 3:1 to about 1:20, about 2:1 to about1:20, about 0:1 to about 1:20, about 1:1 to about 1:20, about 1:2 toabout 1:20, about 1:3 to about 1:20, about 1:4 to about 1:20, about 1:5to about 1:20, about 5:1 to about 1:15, about 4:1 to about 1:15, about3:1 to about 1:15, about 0:1 to about 1:15, about 2:1 to about 1:15,about 1:1 to about 1:15, about 1:2 to about 1:15, about 1:3 to about1:15, about 1:4 to about 1:15, about 1:5 to about 1:15, about 5:1 toabout 1:12, about 4:1 to about 1:12, about 3:1 to about 1:12, about 2:1to about 1:12, about 0:1 to about 1:12, about 1:1 to about 1:12, about1:2 to about 1:12, about 1:3 to about 1:12, about 1:4 to about 1:12,about 1:5 to about 1:12, about 1:6 to about 1:12, about 1:7 to about1:12, about 1:8 to about 1:12, about 5:1 to about 1:10, about 4:1 toabout 1:10, about 3:1 to about 1:10, about 2:1 to about 1:10, about 0:1to about 1:10, about 1:1 to about 1:10, about 1:2 to about 1:10, about1:3 to about 1:10, about 1:4 to about 1:10, about 1:5 to about 1:10,about 1:6 to about 1:10, about 1:7 to about 1:10, or about 1:8 to about1:10.

Step (b) may be carried out at room temperature, in order to allow thesolution comprising the therapeutic compound to form the pharmaceuticalcomposition. However, in other embodiments of the method, step (b) maybe carried out at a temperature that is greater than room temperature,e.g., greater than 21° C., greater than 25° C., greater than 30° C.,greater than 35° C. or greater than 37° C. In certain cases, step (b)may be carried out at temperatures below room temperature, in order toallow a therapeutic compound to dissolve fully in a pharmaceuticallyacceptable solvent. However, in other embodiments of the method, step(b) may be carried out at a temperature that is less than roomtemperature, e.g., less than 10° C., greater than 5° C., greater than 0°C., greater than −10° C. or greater than −20° C. The contacting in step(b) may comprise mixing the solution and the pharmaceutically acceptableadjuvant, e.g., by stirring, inversion, sonication, or vortexing. Themixing may be carried out for, e.g., at least 1 second, at least 5seconds, at least 10 seconds, at least 20 seconds, at least 30 seconds,at least 45 seconds, at least 60 seconds, or more, until thepharmaceutical composition is formed.

In step (c), the solvent removal from a pharmaceutical composition maybe accomplished using one of a variety of procedures known in the art,including, without limitation, evaporation, dialyzation, distillation,lypholization, and filtration. These removal procedures may be doneunder conditions of ambient atmosphere, under low pressure, or under avacuum.

In one embodiment, step (c) may result in the complete removal of apharmaceutically acceptable solvent from the pharmaceutical compositiondisclosed herein. In aspects of this embodiment, step (c) may result in,e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 55%, at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 93%, at least95%, at least 97%, or at least 99% removal of a pharmaceuticallyacceptable solvent from the pharmaceutical composition disclosed herein.

Step (c) is conducted at a temperature that allows for the evaporationof a pharmaceutically acceptable solvent disclosed herein, and as such,an evaporation temperature is solvent-dependent. Factors which influencean evaporation temperature of a solvent include, without limitation, theparticular solvent used, the amount of solvent present, the particulartherapeutic compound present, the particular adjuvant present, thestability of the therapeutic compound present, the reactivity of thetherapeutic compound present, the particular atmospheric pressure used,the time desired for complete evaporation. Generally, a pharmaceuticalcomposition will require heating if the evaporation step is conducted atambient pressure, e.g., 1 atm. However, under high vacuum conditions,the evaporation step may be conducted at temperatures below ambienttemperature, e.g., less than 22° C.

In some aspects, removal of solvent from the pharmaceutical compositionmay be carried out at ambient atmospheric pressure and at a temperatureabove ambient temperature. In aspects of this embodiment, removal ofsolvent from the pharmaceutical composition may be carried out atambient atmospheric pressure and at a temperature of, e.g., more than25° C., more than 30° C., more than 35° C., more than 40° C., more than45° C., more than 50° C., more than 55° C., more than 60° C., more than65° C., more than 70° C., more than 80° C., or more than 85° C. In otheraspects of this embodiment, removal of solvent from the pharmaceuticalcomposition may be carried out at ambient atmospheric pressure and at atemperature in a range of, e.g., about 25° C. to about 100° C., about25° C. to about 95° C., about 25° C. to about 90° C., about 25° C. toabout 85° C., about 25° C. to about 80° C., about 25° C. to about 75°C., about 25° C. to about 70° C., about 25° C. to about 65° C., or about25° C. to about 60° C.

In another embodiment, removal of solvent from the pharmaceuticalcomposition may be carried out under vacuum and at a temperature belowambient temperature. In aspects of this embodiment, removal of solventfrom the pharmaceutical composition may be carried out under vacuum andat a temperature of, e.g., less than 20° C., less than 18° C., less than16° C., less than 14° C., less than 12° C., less than 10° C., less than8° C., less than 6° C., less than 4° C., less than 2° C., or less than0° C. In other aspects of this embodiment, removal of solvent from thepharmaceutical composition may be carried out under vacuum and at atemperature in a range of, e.g., about −20° C. to about 20° C., about−20° C. to about 18° C., about −20° C. to about 16° C., about −20° C. toabout 14° C., about −20° C. to about 12° C., about −20° C. to about 10°C., about −20° C. to about 8° C., about −20° C. to about 6° C., about−20° C. to about 4° C., about −20° C. to about 2° C., about −20° C. toabout 0° C., about −15° C. to about 20° C., about −10° C. to about 20°C., about −5° C. to about 20° C., about 0° C. to about 20° C., about−10° C. to about 20° C., about −10° C. to about 18° C., about −10° C. toabout 16° C., about −10° C. to about 14° C., about −10° C. to about 12°C., about −10° C. to about 10° C., about −10° C. to about 8° C., about−10° C. to about 6° C., about −10° C. to about 4° C., about −10° C. toabout 2° C., or about −10° C. to about 0° C.

The final concentration of a therapeutic compound in a pharmaceuticalcomposition may vary over a wide range and generally may becharacterized as a therapeutically effective amount. In some aspects,the final concentration of a therapeutic compound in a pharmaceuticalcomposition may be, e.g., at least 0.00001 mg/mL, at least 0.0001 mg/mL,at least 0.001 mg/mL, at least 0.01 mg/mL, at least 0.1 mg/mL, at least1 mg/mL, at least 10 mg/mL, at least 25 mg/mL, at least 50 mg/mL, atleast 100 mg/mL, at least 200 mg/mL, at least 500 mg/mL, at least 700mg/mL, at least 1,000 mg/mL, or at least 1,200 mg/mL. In other aspectsof this embodiment, the concentration of a therapeutic compound in thesolution may be, e.g., at most 1,000 mg/mL, at most 1,100 mg/mL, at most1,200 mg/mL, at most 1,300 mg/mL, at most 1,400 mg/mL, at most 1,500mg/mL, at most 2,000 mg/mL, at most 2,000 mg/mL, or at most 3,000 mg/mL.In other aspects of this embodiment, the final concentration of atherapeutic compound in a pharmaceutical composition may be in a rangeof, e.g., about 0.00001 mg/mL to about 3,000 mg/mL, about 0.0001 mg/mLto about 3,000 mg/mL, about 0.01 mg/mL to about 3,000 mg/mL, about 0.1mg/mL to about 3,000 mg/mL, about 1 mg/mL to about 3,000 mg/mL, about250 mg/mL to about 3,000 mg/mL, about 500 mg/mL to about 3,000 mg/mL,about 750 mg/mL to about 3,000 mg/mL, about 1,000 mg/mL to about 3,000mg/mL, about 100 mg/mL to about 2,000 mg/mL, about 250 mg/mL to about2,000 mg/mL, about 500 mg/mL to about 2,000 mg/mL, about 750 mg/mL toabout 2,000 mg/mL, about 1,000 mg/mL to about 2,000 mg/mL, about 100mg/mL to about 1,500 mg/mL, about 250 mg/mL to about 1,500 mg/mL, about500 mg/mL to about 1,500 mg/mL, about 750 mg/mL to about 1,500 mg/mL,about 1,000 mg/mL to about 1,500 mg/mL, about 100 mg/mL to about 1,200mg/mL, about 250 mg/mL to about 1,200 mg/mL, about 500 mg/mL to about1,200 mg/mL, about 750 mg/mL to about 1,200 mg/mL, about 1,000 mg/mL toabout 1,200 mg/mL, about 100 mg/mL to about 1,000 mg/mL, about 250 mg/mLto about 1,000 mg/mL, about 500 mg/mL to about 1,000 mg/mL, about 750mg/mL to about 1,000 mg/mL, about 100 mg/mL to about 750 mg/mL, about250 mg/mL to about 750 mg/mL, about 500 mg/mL to about 750 mg/mL, about100 mg/mL to about 500 mg/mL, about 250 mg/mL to about 500 mg/mL, about0.00001 mg/mL to about 0.0001 mg/mL, about 0.00001 mg/mL to about 0.001mg/mL, about 0.00001 mg/mL to about 0.01 mg/mL, about 0.00001 mg/mL toabout 0.1 mg/mL, about 0.00001 mg/mL to about 1 mg/mL, about 0.001 mg/mLto about 0.01 mg/mL, about 0.001 mg/mL to about 0.1 mg/mL, about 0.001mg/mL to about 1 mg/mL, about 0.001 mg/mL to about 10 mg/mL, or about0.001 mg/mL to about 100 mg/mL.

A pharmaceutical composition produced using the methods disclosed hereinmay be a liquid formulation or a solid or semi-solid formulation. Aliquid formulation can be formed by using various lipids like oils ofother fatty acids that remain as liquids in the temperature rangedesired. In some aspects, a pharmaceutical composition is liquid at roomtemperature. In other aspects, a pharmaceutical composition may beformulated to be a liquid at a temperature of, e.g., about 25° C. orhigher, about 23° C. or higher, about 21° C. or higher, about 19° C. orhigher, about 17° C. or higher, about 15° C. or higher, about 12° C. orhigher, about 10° C. or higher, about 8° C. or higher, about 6° C. orhigher, about 4° C. or higher, or about 0° C. or higher.

A solid or semi-solid formulation may take advantage of the differentmelting point temperatures of the various adjuvants like fatty acids.Formation of a solid or semi-solid dosage form can be by modifying therespective concentrations of the fatty acids comprising a pharmaceuticalcomposition disclosed herein. For example, linolenic acid has a meltingpoint temperature (T_(m)) of about −11° C., linoleic acid has a T_(m) ofabout −5° C., oleic acid has a T_(m) of about 16° C., palmitic acid hasa T_(m) of about 61-62° C., and Stearic acid has a T_(m) of about 67-72°C. Increasing the proportion(s) of palmitic, stearic or oleic acid wouldincrease the overall melting temperature of a composition, while,conversely, increasing the proportion(s) of linoleic and linolenic acidwould decrease the melting temperature of a composition. Thus, bycontrolling the types and amounts of the adjuvant components added, apharmaceutical composition can be made that is substantially solid orsemi-solid at room temperature, but melts when it is ingested, andreaches body temperature. The resulting melted composition readily formsmicelles which are absorbed by the intestine, assembled intochylomicrons, and ultimately absorbed by macrophages. The solid dosageform may be a powder, granule, tablet, capsule or suppository.

Aspects of the present specification disclose a method of treating anindividual with a chronic inflammation, particularly chronicinflammation associated with aging and/or age-related disorders. In oneembodiment, the method comprises the step of administering to anindividual in need thereof a pharmaceutical composition, whereinadministration reduces a symptom associated with the chronicinflammation, thereby treating the individual.

As used herein, the term “treating,” refers to reducing or eliminatingin an individual a clinical symptom of a chronic inflammation; ordelaying or preventing in an individual the onset of a clinical symptomof a chronic inflammation. For example, the term “treating” can meanreducing a symptom of a condition characterized by a chronicinflammation by, e.g., at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 55%, atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90% at least 95%, or at least 100%. The actualsymptoms associated with chronic inflammation are well known and can bedetermined by a person of ordinary skill in the art by taking intoaccount factors, including, without limitation, the location of thechronic inflammation, the cause of the chronic inflammation, theseverity of the chronic inflammation, and/or the tissue or organaffected by the chronic inflammation. Those of skill in the art willknow the appropriate symptoms or indicators associated with a specifictype of chronic inflammation and will know how to determine if anindividual is a candidate for treatment as disclosed herein.

In some aspects, a chronic inflammation comprises a tissue inflammation.Tissue inflammation is a chronic inflammation that is confined to aparticular tissue or organ. In aspect of this embodiment, a tissueinflammation comprises, e.g., a skin inflammation, a muscleinflammation, a tendon inflammation, a ligament inflammation, a boneinflammation, a cartilage inflammation, a lung inflammation, a heartinflammation, a liver inflammation, a pancreatic inflammation, a kidneyinflammation, a bladder inflammation, a stomach inflammation, anintestinal inflammation, a neuron inflammation, and a braininflammation.

In other aspects, a chronic inflammation comprises a systemicinflammation. Although the processes involved are identical to tissueinflammation, systemic inflammation is not confined to a particulartissue but in fact overwhelms the body, involving the endothelium andother organ systems. When it is due to infection, the term sepsis isapplied, with the terms bacteremia being applied specifically forbacterial sepsis and viremia specifically to viral sepsis. Vasodilationand organ dysfunction are serious problems associated with widespreadinfection that may lead to septic shock and death.

In some aspects, a chronic inflammation comprises an arthritis.Arthritis includes a group of conditions involving damage to the jointsof the body due to the inflammation of the synovium including, withoutlimitation osteoarthritis, rheumatoid arthritis, juvenile idiopathicarthritis, spondyloarthropathies like ankylosing spondylitis, reactivearthritis (Reiter's syndrome), psoriatic arthritis, enteropathicarthritis associated with inflammatory bowel disease, Whipple diseaseand Behcet disease, septic arthritis, gout (also known as goutyarthritis, crystal synovitis, metabolic arthritis), pseudogout (calciumpyrophosphate deposition disease), and Still's disease. Arthritis canaffect a single joint (monoarthritis), two to four joints(oligoarthritis) or five or more joints (polyarthritis) and can beeither an auto-immune disease or a non-autoimmune disease.

In some aspects, a chronic inflammation comprises an autoimmunedisorder. Autoimmune diseases can be broadly divided into systemic andorgan-specific autoimmune disorders, depending on the principalclinico-pathologic features of each disease. Systemic autoimmunediseases include, without limitation, systemic lupus erythematosus(SLE), Sjogren's syndrome, Scleroderma, rheumatoid arthritis andpolymyositis. Local autoimmune diseases may be endocrinologic (DiabetesMellitus Type 1, Hashimoto's thyroiditis, Addison's disease etc.),dermatologic (pemphigus vulgaris), hematologic (autoimmune haemolyticanemia), neural (multiple sclerosis) or can involve virtually anycircumscribed mass of body tissue. Types of autoimmune disordersinclude, without limitation, acute disseminated encephalomyelitis(ADEM), Addison's disease, an allergy or sensitivity, amyotrophiclateral sclerosis, anti-phospholipid antibody syndrome (APS), arthritis,autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner eardisease, autoimmune pancreatitis, bullous pemphigoid, celiac disease,Chagas disease, chronic obstructive pulmonary disease (COPD), diabetesmellitus type 1 (IDDM), endometriosis, fibromyalgia, Goodpasture'ssyndrome, Graves' disease, Guillain-Barre syndrome (GBS), Hashimoto'sthyroiditis, hidradenitis suppurativa, idiopathic thrombocytopenicpurpura, inflammatory bowel disease, interstitial cystitis, lupus(including discoid lupus erythematosus, drug-induced lupuserythematosus, lupus nephritis, neonatal lupus, subacute cutaneous lupuserythematosus and systemic lupus erythematosus), morphea, multiplesclerosis (MS), myasthenia gravis, myopathies, narcolepsy,neuromyotonia, pemphigus vulgaris, pernicious anaemia, primary biliarycirrhosis, recurrent disseminated encephalomyelitis (multiphasicdisseminated encephalomyelitis), rheumatic fever, schizophrenia,scleroderma, Sjogren's syndrome, tenosynovitis, vasculitis, andvitiligo.

In another embodiment, a chronic inflammation comprises a myopathy.Myopathies are caused when the immune system inappropriately attackscomponents of the muscle, leading to inflammation in the muscle. Amyopathy includes an inflammatory myopathy and an auto-immune myopathy.Myopathies include, without limitation, dermatomyositis, inclusion bodymyositis, and polymyositis.

A composition or compound as described herein may be administered to anindividual. An individual is typically a human being, but is inclusiveof other warm-blooded animals such as mammals. Typically, any individualwho is a candidate for a conventional chronic inflammation treatment isa candidate for a chronic inflammation treatment disclosed herein.Pre-operative evaluation typically includes routine history and physicalexamination in addition to thorough informed consent disclosing allrelevant risks and benefits of the procedure.

A pharmaceutical composition may comprise a therapeutic compound in atherapeutically effective amount. As used herein, the term “effectiveamount” is synonymous with “therapeutically effective amount,”“effective dose,” or “therapeutically effective dose,” and when used inreference to treating a chronic inflammation refers to the minimum doseof a therapeutic compound necessary to achieve the desired therapeuticeffect and includes a dose sufficient to reduce a symptom associatedwith a chronic inflammation. The effectiveness of a therapeutic compoundin treating a chronic inflammation can be determined by observing animprovement in an individual based upon one or more clinical symptoms,and/or physiological indicators associated with the condition. Animprovement in a chronic inflammation also can be indicated by a reducedneed for a concurrent therapy.

In accordance with one or more aspects disclosed herein, a first dose ofa oxidoreductase inhibitor is administered to an individual. The firstdose may be determined by a person of ordinary skill in the art bytaking into account factors, including, without limitation, the type ofchronic inflammation, the location of the chronic inflammation, thecause of the chronic inflammation, the severity of the chronicinflammation, the degree of relief desired, the duration of reliefdesired, the particular therapeutic compound used, the rate of excretionof the therapeutic compound used, the pharmacodynamics of thetherapeutic compound used, the nature of the other compounds to beincluded in the composition, the particular route of administration, theparticular characteristics, history and risk factors of the patient,such as, e.g., age, weight, general health and the like, or anycombination thereof. Additionally, where repeated administration of atherapeutic compound is used, an effective amount of a therapeuticcompound will further depend upon factors, including, withoutlimitation, the frequency of administration, the half-life of thetherapeutic compound, or any combination thereof. In is known by aperson of ordinary skill in the art that an effective amount of atherapeutic compound can be extrapolated from in vitro assays and invivo administration studies using animal models prior to administrationto humans.

Following the administration of the first dose, levels of nitrite andhydrogen sulfide in the individual may be measured. The levels ofnitrite and hydrogen sulfide may be measured using known techniques,such as by taking a blood sample or urine sample, or using a devicecapable of measuring levels from a probe placed into contact with theskin or other tissue. The measured levels of nitrite and hydrogensulfide may be compared to prescribed ranges. The dose of theoxidoreductase inhibitor(s) thereafter may be adjusted as needed tomaintain nitrite levels and hydrogen sulfide levels within theprescribed ranges. For example, if the measured nitrite level exceedsthe prescribed range and the measured hydrogen sulfide level falls belowthe prescribed range, the dose may be increased by an amountcommensurate with the extent of the excess nitrite and hydrogen sulfidedeficiency, taking the above-noted factors into account. Followingadministration of the adjusted dose, levels of levels of nitrite andhydrogen sulfide may be measured again to determine whether the levelsfall within the prescribed ranges, and subsequent dosing adjustments maybe made thereafter as needed.

In some aspects, a therapeutically effective amount of a therapeuticcompound reduces a symptom associated with a chronic inflammation by,e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least30%, at least 35%, at least 40%, at least 45%, at least 50%, at least55%, at least 60%, at least 65%, at least 70%, at least 75%, at least80%, at least 85%, at least 90%, at least 95% or at least 100%. In otheraspects of this embodiment, a therapeutically effective amount of atherapeutic compound reduces a symptom associated with a chronicinflammation by, e.g., at most 10%, at most 15%, at most 20%, at most25%, at most 30%, at most 35%, at most 40%, at most 45%, at most 50%, atmost 55%, at most 60%, at most 65%, at most 70%, at most 75%, at most80%, at most 85%, at most 90%, at most 95% or at most 100%. In yet otheraspects of this embodiment, a therapeutically effective amount of atherapeutic compound reduces a symptom associated with a chronicinflammation by, e.g., about 10% to about 100%, about 10% to about 90%,about 10% to about 80%, about 10% to about 70%, about 10% to about 60%,about 10% to about 50%, about 10% to about 40%, about 20% to about 100%,about 20% to about 90%, about 20% to about 80%, about 20% to about 20%,about 20% to about 60%, about 20% to about 50%, about 20% to about 40%,about 30% to about 100%, about 30% to about 90%, about 30% to about 80%,about 30% to about 70%, about 30% to about 60%, or about 30% to about50%.

In some aspects, a therapeutically effective amount of a therapeuticcompound generally is in the range of about 0.001 mg/kg/day to about 100mg/kg/day. In aspects of this embodiment, an effective amount of atherapeutic compound may be, e.g., at least 0.001 mg/kg/day, at least0.01 mg/kg/day, at least 0.1 mg/kg/day, at least 1.0 mg/kg/day, at least5.0 mg/kg/day, at least 10 mg/kg/day, at least 15 mg/kg/day, at least 20mg/kg/day, at least 25 mg/kg/day, at least 30 mg/kg/day, at least 35mg/kg/day, at least 40 mg/kg/day, at least 45 mg/kg/day, or at least 50mg/kg/day. In other aspects of this embodiment, an effective amount of atherapeutic compound may be in the range of, e.g., about 0.001 mg/kg/dayto about 10 mg/kg/day, about 0.001 mg/kg/day to about 15 mg/kg/day,about 0.001 mg/kg/day to about 20 mg/kg/day, about 0.001 mg/kg/day toabout 25 mg/kg/day, about 0.001 mg/kg/day to about 30 mg/kg/day, about0.001 mg/kg/day to about 35 mg/kg/day, about 0.001 mg/kg/day to about 40mg/kg/day, about 0.001 mg/kg/day to about 45 mg/kg/day, about 0.001mg/kg/day to about 50 mg/kg/day, about 0.001 mg/kg/day to about 75mg/kg/day, or about 0.001 mg/kg/day to about 100 mg/kg/day. In yet otheraspects of this embodiment, an effective amount of a therapeuticcompound may be in the range of, e.g., about 0.01 mg/kg/day to about 10mg/kg/day, about 0.01 mg/kg/day to about 15 mg/kg/day, about 0.01mg/kg/day to about 20 mg/kg/day, about 0.01 mg/kg/day to about 25mg/kg/day, about 0.01 mg/kg/day to about 30 mg/kg/day, about 0.01mg/kg/day to about 35 mg/kg/day, about 0.01 mg/kg/day to about 40mg/kg/day, about 0.01 mg/kg/day to about 45 mg/kg/day, about 0.01mg/kg/day to about 50 mg/kg/day, about 0.01 mg/kg/day to about 75mg/kg/day, or about 0.01 mg/kg/day to about 100 mg/kg/day. In stillother aspects of this embodiment, an effective amount of a therapeuticcompound may be in the range of, e.g., about 0.1 mg/kg/day to about 10mg/kg/day, about 0.1 mg/kg/day to about 15 mg/kg/day, about 0.1mg/kg/day to about 20 mg/kg/day, about 0.1 mg/kg/day to about 25mg/kg/day, about 0.1 mg/kg/day to about 30 mg/kg/day, about 0.1mg/kg/day to about 35 mg/kg/day, about 0.1 mg/kg/day to about 40mg/kg/day, about 0.1 mg/kg/day to about 45 mg/kg/day, about 0.1mg/kg/day to about 50 mg/kg/day, about 0.1 mg/kg/day to about 75mg/kg/day, or about 0.1 mg/kg/day to about 100 mg/kg/day.

In other aspects of this embodiment, an effective amount of atherapeutic compound may be in the range of, e.g., about 1 mg/kg/day toabout 10 mg/kg/day, about 1 mg/kg/day to about 15 mg/kg/day, about 1mg/kg/day to about 20 mg/kg/day, about 1 mg/kg/day to about 25mg/kg/day, about 1 mg/kg/day to about 30 mg/kg/day, about 1 mg/kg/day toabout 35 mg/kg/day, about 1 mg/kg/day to about 40 mg/kg/day, about 1mg/kg/day to about 45 mg/kg/day, about 1 mg/kg/day to about 50mg/kg/day, about 1 mg/kg/day to about 75 mg/kg/day, or about 1 mg/kg/dayto about 100 mg/kg/day. In yet other aspects of this embodiment, aneffective amount of a therapeutic compound may be in the range of, e.g.,about 5 mg/kg/day to about 10 mg/kg/day, about 5 mg/kg/day to about 15mg/kg/day, about 5 mg/kg/day to about 20 mg/kg/day, about 5 mg/kg/day toabout 25 mg/kg/day, about 5 mg/kg/day to about 30 mg/kg/day, about 5mg/kg/day to about 35 mg/kg/day, about 5 mg/kg/day to about 40mg/kg/day, about 5 mg/kg/day to about 45 mg/kg/day, about 5 mg/kg/day toabout 50 mg/kg/day, about 5 mg/kg/day to about 75 mg/kg/day, or about 5mg/kg/day to about 100 mg/kg/day.

Dosing can be single dosage or cumulative (serial dosing), and can bereadily determined by one skilled in the art. For instance, treatment ofa chronic inflammation may comprise a one-time administration of aneffective dose of a pharmaceutical composition disclosed herein.Alternatively, treatment of a chronic inflammation may comprise multipleadministrations of an effective dose of a pharmaceutical compositioncarried out over a range of time periods, such as, e.g., once daily,twice daily, trice daily, once every few days, or once weekly. Thetiming of administration can vary from individual to individual,depending upon such factors as the severity of an individual's symptoms.For example, an effective dose of a pharmaceutical composition can beadministered to an individual once daily for an indefinite period oftime, or until the individual no longer requires therapy. A person ofordinary skill in the art will recognize that the condition of theindividual can be monitored throughout the course of treatment and thatthe effective amount of a pharmaceutical composition that isadministered can be adjusted accordingly.

In one embodiment, upon administration to an individual, apharmaceutical composition comprising a therapeutic compound results ina bio-distribution of the therapeutic compound different than abio-distribution of the therapeutic compound included in the samepharmaceutical composition, except without an adjuvant disclosed herein.

In another embodiment, upon administration to an individual, atherapeutic compound of the pharmaceutical composition is delivered to amacrophage. Macrophages are one of the key cell types believed to beinvolved in the control of the inflammation response. The resultant highlevel of a therapeutic compound having anti-inflammatory activitypresent in the macrophages results in a clinically effective treatmentof chronic inflammation. In an aspect of this embodiment, uponadministration to an individual, a therapeutically effective amount of atherapeutic compound of the pharmaceutical composition is preferentiallydelivered to a macrophage. In other aspect of this embodiment, uponadministration to an individual, a therapeutic compound of thepharmaceutical composition is substantially delivered to a macrophage.In yet other aspect of this embodiment, upon administration to anindividual, the amount of a therapeutic compound of the pharmaceuticalcomposition delivered to a macrophage is, e.g., at least 5%, at least10%, at least 15%, at least 20%, at least 25%, at least 30%, at least35%, at least 40%, at least 45%, at least 50%, at least 55%, at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, or at least 100% of the total amount ofthe therapeutic compound contained in the administered pharmaceuticalcomposition. In still other aspects of this embodiment, uponadministration to an individual, the amount of a therapeutic compound ofthe pharmaceutical composition delivered to a macrophage is in a rangeof, e.g., about 5% to about 100%, about 10% to about 100%, about 15% toabout 100%, about 20% to about 100%, about 25% to about 100%, about 30%to about 100%, about 35% to about 100%, about 40% to about 100%, about45% to about 100%, about 50% to about 100%, about 5% to about 90%, about10% to about 90%, about 15% to about 90%, about 20% to about 90%, about25% to about 90%, about 30% to about 90%, about 35% to about 90%, about40% to about 90%, about 45% to about 90%, about 50% to about 90%, about5% to about 80%, about 10% to about 80%, about 15% to about 80%, about20% to about 80%, about 25% to about 80%, about 30% to about 80%, about35% to about 80%, about 40% to about 80%, about 45% to about 80%, about50% to about 80%, about 5% to about 70%, about 10% to about 70%, about15% to about 70%, about 20% to about 70%, about 25% to about 70%, about30% to about 70%, about 35% to about 70%, about 40% to about 70%, about45% to about 70%, or about 50% to about 70% of the total amount of thetherapeutic compound contained in the administered pharmaceuticalcomposition.

A pharmaceutical composition can also be administered to an individualin combination with other therapeutic compounds to increase the overalltherapeutic effect of the treatment. The use of multiple compounds totreat an indication can increase the beneficial effects while reducingthe presence of side effects.

The following examples illustrate but do not limit the scope of thedisclosure as set forth herein.

Example 1

This example describes an experiment to determine whether treatmentswith isomyosmine have an effect on hydrogen sulfide (H₂S) quantity inmice with experimental autoimmune encephalomyelitis (EAE). Mice wereimmunized for EAE on day 0 and treated with either vehicle control or 5mg/ml isomyosmine in water, for the duration of the experiment. Bloodwas collected, serum was separated and assayed for H₂S via KamiyaBiomedical ELISA kit per the manufacturer's instructions. Two differentexperiments were performed. In the first experiment, blood was collectedat the end of disease course. In the second experiment, blood wascollected at day 12 after immunization. In both cases, as shown FIGS. 1Aand 1B, mice treated with isomyosmine had a trend toward an increase inH₂S as measured by ELISA.

Example 2

This example describes an experiment to test whether isomyosminedirectly inhibits nitrate reductase in a cell-free setting. Standardconcentrations of nitrate were incubated with purified nitrate reductaseand vehicle or isomyosmine, and the Griess reaction was used to measurenitrite. Several concentrations of nitrate and two concentrations ofisomyosmine were used. Isomyosmine was found to inhibit nitratereductase, most efficiently with a low amount of substrate (FIG. 2A) andto a lesser extent with a medium amount of substrate (FIG. 2B). Once thesystem becomes saturated (high amount of substrate), the inhibitoryeffect was reduced (FIG. 2C). These data suggest that the decrease is atleast partly the result of a direct effect of isomyosmine on the nitratereductase itself.

Example 3

This example describes an experiment that may be used for the detectionand quantitation of protein carbonyls (FIG. 3). Protein Carbonyl ELISAKit (ABIN2344951 from Cell Biolabs Inc., San Diego, Calif.) is an enzymeimmunoassay developed for this purpose. The protein carbonyls present ina sample (or standard) are derivatized to DNP hydrazone and probed withan anti-DNP antibody, followed by an HRP conjugated secondary antibody.The protein carbonyl content in an unknown sample may be determined bycomparing with a standard curve that is prepared from predeterminedreduced and oxidized BSA standards.

Example 4

This example describes an assay which may be used for determiningxanthine oxidase activity (FIG. 4). Xanthine oxidase is present inappreciable amounts in liver and jejunum in normal conditions. However,in various liver disorders and in inflammatory conditions, XO isreleased into circulation. Therefore, we hypothesize that the serum XOdetermination could be an indicator of the chronic, sterile, low-gradeinflammation develops, which contributes to the pathogenesis ofage-related diseases (phenomenon so-called inflammaging). Franceschi,“Inflammaging: a new immune-metabolic viewpoint for age-relateddiseases,” Nature Reviews Endocrinology, 2018, vol. 14, 576-590.

Xanthine Oxidase Activity Assay Kit (ab102522 from Abcam, Cambridge,Mass.) is a colorimetric/fluorometric assay which may be used todetermine xanthine oxidase activity in variety of samples. In thexanthine oxidase assay protocol, xanthine oxidase oxidizes xanthine tohydrogen peroxide (H₂O₂) which reacts stoichiometrically with a probe togenerate color (at OD=570 nm) and fluorescence (at Ex/Em=535/587 nm).Since the color or fluorescence intensity is proportional to XO content,the XO activity can be accurately measured.

While particular embodiments have been described and illustrated, itshould be understood that the invention is not limited thereto sincemodifications may be made by persons skilled in the art. The presentapplication contemplates any and all modifications that fall within thespirit and scope of the underlying invention disclosed and claimedherein.

1. A method of regulating oxidoreductase activity for treating aninflammation or age-related disorder comprising administering to anindividual in need thereof a therapeutically effective amount ofisomyosmine or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable vehicle therefor.
 2. The method of claim 1,wherein the therapeutically effective amount is from about 0.001 toabout 75 mg/kg/day.
 3. The method of claim 1, wherein thetherapeutically effective amount is from about 0.01 to about 50mg/kg/day.
 4. The method of claim 1, wherein the therapeuticallyeffective amount is from about 0.1 to about 40 mg/kg/day.
 5. The methodof claim 1, wherein the therapeutically effective amount is from about 1to about 30 mg/kg/day.
 6. The method of claim 1, wherein thetherapeutically effective amount is from about 1 to about 20 mg/kg/day.7. The method of claim 1, wherein the therapeutically effective amountis from about 1 to about 10 mg/kg/day.
 8. A method of treating oxidativestress associated with an inflammation or age-related disordercomprising administering to an individual in need thereof atherapeutically effective amount of isomyosmine or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable vehicletherefor.
 9. The method of claim 8, wherein the therapeuticallyeffective amount is from about 0.001 to about 75 mg/kg/day.
 10. Themethod of claim 8, wherein the therapeutically effective amount is fromabout 0.01 to about 50 mg/kg/day.
 11. The method of claim 8, wherein thetherapeutically effective amount is from about 0.1 to about 40mg/kg/day.
 12. The method of claim 8, wherein the therapeuticallyeffective amount is from about 1 to about 30 mg/kg/day.
 13. The methodof claim 8, wherein the therapeutically effective amount is from about 1to about 20 mg/kg/day.
 14. The method of claim 8, wherein thetherapeutically effective amount is from about 1 to about 10 mg/kg/day.15. A method of treating a disorder comprising administering to anindividual in need thereof a therapeutically effective amount ofisomyosmine or a pharmaceutically acceptable salt thereof, where thedisorder is selected from the group consisting of infectious orparasitic diseases; neoplasms; diseases of the blood or blood-formingorgans; diseases of the immune system; endocrine; nutritional ormetabolic diseases; mental, behavioral or neurodevelopmental disorders;sleep-wake disorders; diseases of the nervous system; diseases of thevisual system; diseases of the ear or mastoid process; diseases of thecirculatory system; diseases of the respiratory system; diseases of thedigestive system; diseases of the skin; diseases of the musculoskeletalsystem or connective tissue; diseases of the genitourinary system;conditions related to sexual health; pregnancy, childbirth or thepuerperium; conditions originating in the perinatal period;developmental anomalies; and injury, poisoning or other consequences ofexternal causes.
 16. The method of claim 15, wherein the disorder ismalaria.
 17. The method of claim 15, wherein the disorder is Lymedisease.
 18. The method of claim 15, wherein the disorder is anxiety.19. The method of claim 15, wherein the disorder is depression.
 20. Themethod of claim 15, wherein the disorder is an age-related disease ofthe musculoskeletal system.